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Journal articles on the topic 'Hybridoma's'

Author: Grafiati

Published: 4 June 2021

Last updated: 29 July 2025

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1

Ozaki, S., S. K. Durum, K. Muegge, J. York-Jolley, and J. A. Berzofsky. "Production of T-T hybrids from T cell clones. Direct comparison between cloned T cells and T hybridoma cells derived from them." Journal of Immunology 141, no. 1 (1988): 71–78. http://dx.doi.org/10.4049/jimmunol.141.1.71.

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Abstract It has been assumed, without direct evidence, that T cell hybridomas and non-transformed T cell clones are both good models of normal Ag-specific T cells. To compare directly the difference in activation of cloned normal T cells and T hybridoma cells with the same TCR, cloned T hybridoma cells were obtained by fusing pre-established, myoglobin-specific, Iad-restricted T cell clones (14.5 and 9.27) with BW5147 cells. T cell clones were pre-activated with IL-2 as well as specific Ag before fusion. Cloned T hybridoma A3.4C6 was derived from Lys 140-specific and I-Ed-restricted clone 14.5

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2

Hawksworth, David, Jeff Moore, and Isaac Larkin. "Diversity and affinity from plasma vs. B cells for monoclonal antibody development." Journal of Immunology 212, no. 1_Supplement (2024): 0234_4543. http://dx.doi.org/10.4049/jimmunol.212.supp.0234.4543.

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Abstract Diagnostic assay development requires a diverse panel of high affinity antibodies. Historically, our lab has used splenic CD19+ B-cells to generate mouse hybridoma cell lines. However, literature suggests that, as a group, terminally differentiated, CD138+ plasma cells secrete the highest affinity antibodies, making these cells an attractive target for hybridoma development. To compare the use of these two lymphocyte populations for generating mouse hybridomas, three fusion experiments were completed whereby cells were separately isolated and fused. Hybridomas were plated in semi-soli

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3

Rauch, J., H. Massicotte, and H. Tannenbaum. "Hybridoma anti-DNA autoantibodies from patients with rheumatoid arthritis and systemic lupus erythematosus demonstrate similar nucleic acid binding characteristics." Journal of Immunology 134, no. 1 (1985): 180–86. http://dx.doi.org/10.4049/jimmunol.134.1.180.

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Abstract Hybridoma anti-DNA antibodies have been generated from the fusion of the GM 4672 lymphoblastoid line with peripheral blood lymphocytes from four normal subjects, nine patients with rheumatoid arthritis (RA), and 13 patients with systemic lupus erythematosus (SLE). A total of 441 hybridoma clones were obtained, of which 37 secreted anti-DNA autoantibodies. The nucleic acid binding characteristics of the anti-DNA antibodies produced by two hybridomas from normal subjects, nine hybridomas from RA patients, and 18 hybridomas from SLE patients are reported. The hybridoma anti-DNA antibodie

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4

Kuchroo, V. K., P. R. Billings, C. Levine, C. A. Martin, R. T. Kubo, and M. E. Dorf. "Antigen-specific and antibody-mediated growth inhibition of suppressor T cell hybridomas. Roles of H-2 and the CD3-T cell receptor-alpha/beta complex." Journal of Immunology 145, no. 4 (1990): 1059–65. http://dx.doi.org/10.4049/jimmunol.145.4.1059.

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Abstract Eight different Ts cell hybridomas (including inducer (Ts1) and effector (Ts3) suppressor cells) specific for the 4-hydroxy-3-nitrophenyl acetyl (NP) hapten were tested for their ability to respond to Ag or anti-CD3 antibody in a growth-inhibition assay. Results suggest that the expression of the TCR-CD3 complex on Ts hybridomas is required for the Ag or anti-CD3-mediated growth inhibition. One of the CD3+, Ts hybridomas (CKB-Ts3-9.H3) was tested in detail; this CD4- effector suppressor cell hybridoma showed specific inhibition of growth in the presence of NP or NIP-coupled protein co

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5

Machida, Keigo, Yasuteru Kondo, Jeffrey Y. Huang, et al. "Hepatitis C Virus (HCV)-Induced Immunoglobulin Hypermutation Reduces the Affinity and Neutralizing Activities of Antibodies against HCV Envelope Protein." Journal of Virology 82, no. 13 (2008): 6711–20. http://dx.doi.org/10.1128/jvi.02582-07.

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ABSTRACT Hepatitis C virus (HCV) often causes persistent infection despite the presence of neutralizing antibodies against the virus in the sera of hepatitis C patients. HCV infects both hepatocytes and B cells through the binding of its envelope glycoprotein E2 to CD81, the putative viral receptor. Previously, we have shown that E2-CD81 interaction induces hypermutation of heavy-chain immunoglobulin (V H ) in B cells. We hypothesize that if HCV infects antibody-producing B cells, the resultant hypermutation of VH may lower the affinity and specificity of the HCV-specific antibodies, enabling

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6

Retter, M. W., R. A. Eisenberg, P. L. Cohen, and S. H. Clarke. "Sm and DNA binding by dual reactive B cells requires distinct VH, V kappa, and VH CDR3 structures." Journal of Immunology 155, no. 4 (1995): 2248–57. http://dx.doi.org/10.4049/jimmunol.155.4.2248.

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Abstract We have previously demonstrated an overlap of the anti-Sm and anti-DNA responses in MRL/Mp-lpr/lpr mice. The Ab produced by many anti-Sm hybridomas bind DNA and are encoded by Ig V genes used by anti-DNA hybridomas. In addition, some anti-Sm Ab that bind DNA have acquired mutations that improve DNA binding, indicating that DNA is a selecting Ag in the anti-Sm response. To gain insight into the basis for the dual binding ability of these Ab, we coexpressed the H chain from the anti-Sm hybridoma 2-12 with nine different L chains. Hybridoma 2-12 binds Sm but not DNA, yet expresses the sa

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7

Tiebout, R. F., F. van Boxtel-Oosterhof, E. A. Stricker, and W. P. Zeijlemaker. "A human hybrid hybridoma." Journal of Immunology 139, no. 10 (1987): 3402–5. http://dx.doi.org/10.4049/jimmunol.139.10.3402.

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Abstract Hybrid hybridomas are obtained by fusion of two cells, each producing its own antibody. Several authors have reported the construction of murine hybrid hybridomas with the aim to obtain bispecific monoclonal antibodies. We have investigated, in a model system, the feasibility of constructing a human hybrid hybridoma. We fused two monoclonal cell lines: an ouabain-sensitive and azaserine/hypoxanthine-resistant Epstein-Barr virus-transformed human cell line that produces an IgG1 kappa antibody directed against tetanus toxoid and an azaserine/hypoxanthine-sensitive and ouabain-resistant

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8

Rath, S., J. Durdik, R. M. Gerstein, E. Selsing, and A. Nisonoff. "Quantitative analysis of idiotypic mimicry and allelic exclusion in mice with a mu Ig transgene." Journal of Immunology 143, no. 6 (1989): 2074–80. http://dx.doi.org/10.4049/jimmunol.143.6.2074.

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Abstract Analysis of C57BL/6 mice (IgM allotype, Igh-6b or mu b) that carry an Ig H chain transgene of a different allotype (mu a) shows that IgM molecules of mixed allotype (mu a mu b) are present among serum antibodies. The finding was extended to hybridomas prepared from nonimmune transgenic mice, many of which also failed to exhibit allelic exclusion. The proportions of mu a and mu b secreted by individual hybridomas varied markedly, and the product of an individual hybridoma was found to be heterogeneous with respect to the allotype content of individual molecules. The ratio of mu a:mu b

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9

Mathur, A., B. G. Van Ness, and R. G. Lynch. "In vivo and in vitro regulation of IgE production in murine hybridomas." Journal of Immunology 145, no. 11 (1990): 3610–17. http://dx.doi.org/10.4049/jimmunol.145.11.3610.

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Abstract Normal BALB/c mice injected i.p. with the IgE-secreting hybridomas B53 (epsilon, kappa anti-DNP), SE1.3 (epsilon, kappa, anti-arsonate) or A3B1 (epsilon, kappa, anti-TNP) were monitored for serum IgE concentrations and frequencies of splenic T lymphocytes with surface membrane receptors for the Fc portion of IgE (Fc epsilon R+ T lymphocytes). Mice with B53 or SE1.3 hybridomas initially developed high concentrations of IgE and CD8+ Fc epsilon R+ T lymphocytes, followed by a progressive decline in both serum IgE and expression of cytoplasmic epsilon-chains in the hybridoma cells. Serum

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10

Kawasaki, H., C. A. Martin, T. Uchida, et al. "Functional analysis of cloned macrophage hybridomas. V. Induction of suppressor T cell responses." Journal of Immunology 137, no. 7 (1986): 2145–51. http://dx.doi.org/10.4049/jimmunol.137.7.2145.

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Abstract It has been suggested that macrophage-like accessory cells are involved in suppressor T cell (Ts) induction. To further analyze this issue, we obtained several cloned macrophage hybridoma cell lines by somatic cell fusion of the macrophage tumor P388D1 of DBA/2 (H-2d) origin with splenic adherent cells of CKB mice (H-2k). Several cloned lines displayed the serological and functional characteristics of macrophages. We evaluated the ability of these hybridomas to induce third order or effector Ts (Ts3) to suppress the contact sensitivity response against the hapten 4-hydroxy-3-nitrophen

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11

Kulczycki, A., J. Trial, J. M. Connolly, S. Sharp, and J. A. Kapp. "Structure and expression of Fc gamma receptors on mouse suppressor T cell hybridomas." Journal of Immunology 137, no. 7 (1986): 2325–30. http://dx.doi.org/10.4049/jimmunol.137.7.2325.

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Abstract Antigen-specific and idiotype-specific mouse suppressor T cell hybridomas were analyzed for the presence and specificity of Fc gamma receptors (Fc gamma R) by EA rosetting and by flow microfluorometry with the use of monoclonal antibodies. We found that four hybridomas expressed Fc gamma R specific for IgG1 and IgG2b, one of which became Fc gamma R- during prolonged culture. Four other hybridomas and the fusion parent, BW5147, consistently lacked Fc gamma R. The 125I-labeled Fc gamma R were isolated from surface radioiodinated hybridoma cells solubilized with 1% Nonidet P-40, were pur

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12

March, A. Kennel-De, C. Prin-Mathieu, C. H. Kohler, M. N. Kolopp-Sarda, G. C. Faure, and M. C. Béné. "Back-Pack Mice as a Model of Renal Mesangial IgA Dimers Deposition." International Journal of Immunopathology and Pharmacology 18, no. 4 (2005): 701–8. http://dx.doi.org/10.1177/039463200501800412.

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Mesangial IgA in IgA nephropathy are dimers with a J chain but no poly-Ig receptor. This molecular structure has led to the hypothesis that these IgA are issued from the lamina propria of mucosal areas, reaching the kidney by way of the peripheral blood. The availability of hybridomas producing IgA dimers provided an opportunity to test this hypothesis in a new experimental model of IgA nephropathy. Mice were injected subcutaneously (back-pack mice) or intraperitoneally with hybridoma cells secreting either monoclonal IgA dimers, or monoclonal IgA monomers. The influence of immune complex form

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13

Iwata, M., M. Adachi, and K. Ishizaka. "Antigen-specific T cells that form IgE-potentiating factor, IgG-potentiating factor, and antigen-specific glycosylation-enhancing factor on antigenic stimulation." Journal of Immunology 140, no. 8 (1988): 2534–42. http://dx.doi.org/10.4049/jimmunol.140.8.2534.

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Abstract BDF1 mice were immunized with alum-absorbed OVA and T cell hybridomas were constructed from their splenic T cells. Many of the hybridomas constitutively produced glycosylation enhancing factor (GEF), which could switch the T cell hybridoma 23A4 cells from the formation of IgE-suppressive factors to the formation of IgE-potentiating factors. When one of the hybridoma clones, 12H5, was incubated with OVA-pulsed syngeneic or semi-syngeneic (H-2b) macrophages, the hybridoma produced GEF that have affinity for OVA, but not for either keyhole limpet hemocyanin or BSA. However, the same hybr

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14

Lanier, L. L., J. J. Ruitenberg, and J. H. Phillips. "Functional and biochemical analysis of CD16 antigen on natural killer cells and granulocytes." Journal of Immunology 141, no. 10 (1988): 3478–85. http://dx.doi.org/10.4049/jimmunol.141.10.3478.

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Abstract CD16 Ag is associated with the low affinity FcR for IgG expressed on human NK cells and granulocytes. In this study, we demonstrate that NK cells specifically lyse murine anti-CD16 hybridoma cell lines, but do not lyse hybridomas against other cell surface differentiation Ag expressed on NK cells. Moreover, the CD18 structure is involved in the CD16-specific xenogeneic interaction between human effector cells and murine hybridoma target cells. Although interaction with anti-CD16 hybridomas or antibodies triggers the cytolytic mechanism of NK cells, this interaction does not induce cel

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15

Iwata, M., K. Katamura, R. T. Kubo, and K. Ishizaka. "Relationship between T cell receptors and antigen-binding factors. I. Specificity of functional T cell receptors on mouse T cell hybridomas that produce antigen-binding T cell factors." Journal of Immunology 143, no. 12 (1989): 3909–16. http://dx.doi.org/10.4049/jimmunol.143.12.3909.

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Abstract Upon antigenic stimulation with OVA-pulsed syngeneic macrophages, the mouse T cell hybridoma 231F1 produced glycosylation inhibiting factor (GIF) having affinity for OVA and IgE-suppressive factors, whereas another T cell hybridoma, 12H5, cells produced OVA-binding glycosylation enhancing factor (GEF) and IgE-potentiating factor. The OVA-binding GIF from the 231F1 cells is an Ag-specific Ts cell factor, whereas OVA-binding GEF from the 12H5 cells is an Ag-specific augmenting factor. Both hybridomas express CD3 complex and functional TCR-alpha beta. Cross-linking of TCR-alpha beta or C

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16

Dersimonian, H., R. S. Schwartz, K. J. Barrett, and B. D. Stollar. "Relationship of human variable region heavy chain germ-line genes to genes encoding anti-DNA autoantibodies." Journal of Immunology 139, no. 7 (1987): 2496–501. http://dx.doi.org/10.4049/jimmunol.139.7.2496.

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Abstract In order to identify the V region genes encoding systemic lupus erythematosus (SLE)-derived anti-DNA autoantibodies, we have determined the nucleotide sequence of heavy chain mRNA from several DNA-binding immunoglobulins secreted by human hybridomas. We used the technique of cDNA primer extension for determining sequences of the VH, D, and JH gene segments of anti-DNA autoantibodies from three different primary hybridoma growths from an SLE patient and one hybridoma from a leprosy patient. Immunoglobulins from two of the SLE hybridomas expressed the same idiotype, Id-16/6, which is al

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17

Gorczynski, R. M., Z. Chen, H. Zeng, and X. M. Fu. "Specificity for in vivo graft prolongation in gamma delta T cell receptor+ hybridomas derived from mice given portal vein donor-specific preimmunization and skin allografts." Journal of Immunology 159, no. 8 (1997): 3698–706. http://dx.doi.org/10.4049/jimmunol.159.8.3698.

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Abstract gamma delta TCR+ hybridoma cells prepared from mesenteric lymph node cells of animals receiving donor-specific immunization via the portal vein can adoptively transfer this increased graft survival to naive animals. Analysis of TCR gamma-chain junctional sequence diversity suggested that some 40 to 50% of the hybridomas expressed gamma-chain junctional sequence diversity and were stimulated to produce cytokines both by heat shock proteins and by minor histocompatibility Ag-specific irradiated peritoneal cells. The remaining gamma delta TCR+ hybridoma cells expressed TCR with a common

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18

Hurwitz, J. L., J. Samaridis, and J. Pelkonen. "Immature and advanced patterns of T cell receptor gene rearrangement among lymphocytes in splenic culture." Journal of Immunology 142, no. 7 (1989): 2533–39. http://dx.doi.org/10.4049/jimmunol.142.7.2533.

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Abstract Bulk populations and 39 hybridomas from splenic Con A cultures were analyzed for rearrangements among TCR genes: alpha, beta, gamma, and delta. Patterns were categorized to reveal general rules governing gene rearrangement within the activated adult peripheral population. Many patterns of gene rearrangement were consistent with previous studies of T cell lines. Additional points of interest were the following: 1) A large proportion of Con A-stimulated splenic cells bore no TCR gene rearrangements. 2) One splenic hybridoma exhibited an unusual gene pattern, with rearrangements, at alph

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19

Bergman, M. C., J. F. Attrep, A. C. Grammer, and P. E. Lipsky. "Ligation of CD40 influences the function of human Ig-secreting B cell hybridomas both positively and negatively." Journal of Immunology 156, no. 9 (1996): 3118–32. http://dx.doi.org/10.4049/jimmunol.156.9.3118.

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Abstract The effect of ligation of CD40 on the proliferation and Ig secretion of a battery of human Ig-secreting hybridomas was examined to determine the regulatory activity of this surface molecule on B cells after initial activation. B cell hybridomas were generated by fusing activated peripheral blood B cells with SPAZ-4, a non-Ig-secreting fusion partner, and were cloned before analysis. All hybridomas expressed CD40 comparably. These hybridomas were stimulated with either recombinant baculovirus-expressed membrane-bound CD40L or a soluble murine CD40L/CD8 construct in the presence or the

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20

Urnovitz, H. B., Y. Chang, M. Scott, J. Fleischman, and R. G. Lynch. "IgA:IgM and IgA:IgA hybrid hybridomas secrete heteropolymeric immunoglobulins that are polyvalent and bispecific." Journal of Immunology 140, no. 2 (1988): 558–63. http://dx.doi.org/10.4049/jimmunol.140.2.558.

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Abstract Polyvalent bispecific antibodies were secreted by hybrid hybridoma cells when both parental clones expressed a naturally polymerizing immunoglobulin. Hybrid hybridomas made from IgA lambda 2 anti-trinitrophenyl (TNP) and IgA kappa anti-phosphocholine (PC) parental cells secreted polymeric IgA antibodies that bound both TNP and PC. Some of the TNP binding was dissociated from the PC binding under conditions of mild reduction and alkylation suggesting that the bispecific polymeric IgA contained disulfide-linked parental monomers as well as bispecific hybrid monomers. Hybrid hybridomas c

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21

Kisaki, T., T. F. Huff, D. H. Conrad, J. Yodoi, and K. Ishizaka. "Monoclonal antibody specific for T cell-derived human IgE binding factors." Journal of Immunology 138, no. 10 (1987): 3345–51. http://dx.doi.org/10.4049/jimmunol.138.10.3345.

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Abstract A B cell hybridoma secreting monoclonal antibody against human IgE binding factors was obtained by immunization of BALB/c mice with partially purified IgE binding factors, and fusion of their spleen cells with SP-2/0-AG14 cells. The monoclonal antibody bound all of the 60,000, 30,000, and 15,000 dalton IgE binding factors from two T cell hybridomas and those from activated T cells of a normal individual. The antibody bound both IgE-potentiating factors, which had affinity for lentil lectin, and IgE-suppressive factors, which had affinity for peanut agglutinin. However, the monoclonal

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22

Blanckmeister, C. A., K. Yamamoto, M. M. Davis, and G. J. Hämmerling. "Antigen-specific, I-A-restricted suppressor hybridomas with spontaneous cytolytic activity. Functional properties and lack of rearrangement of the T cell receptor beta chain genes." Journal of Experimental Medicine 162, no. 3 (1985): 851–63. http://dx.doi.org/10.1084/jem.162.3.851.

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T suppressor (Ts) hybridomas were produced by fusion of the III/4 T cell hybridoma with splenic T cells from CBA mice tolerized with subimmunogenic doses of bovine serum albumin (BSA). Both the Ts hybridoma cells and a suppressor factor (TsF) inhibited in an antigen-specific and I-Ak-restricted fashion the in vitro proliferative response of BSA-immunized lymph node cells. In addition to the suppressive activity, the hybridoma lines displayed spontaneous cytotoxicity against various tumor targets. The isolation of Ts subclones that are suppressive but not cytolytic, as well as the existence of

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Kuklina, G. V., G. D. Elagin, D. V. Pechenkin, et al. "Manufacturing of hybridomas, producing monoclonal antibodies against Burkholderia mallei and Burkholderia pseudomallei antigens." Journal of microbiology epidemiology immunobiology, no. 4 (September 2, 2019): 78–82. http://dx.doi.org/10.36233/0372-9311-2019-4-78-82.

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Aim. Obtaining hybridomas, stable producing specific monoclonal antibodies against Burkholderia mallei and Burkholderia pseudomallei antigens. Materials and methods. The microbial cultures from State Collection of Microorganisms from the Branch of 48 CSRI of the Defense Ministry of Russian Federation (Kirov) and BALB/c mouse were used in research. Hybridization of B lymphocytes with SP2/0-Ag14 myeloma cells was performed by G.Kohler and C.Milstein procedure in De St. Fazekas and D.Scheidegger modification. The specific activity of immune sera, hybridoma supernatants, ascites and evaluating the

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Takahashi, M., and S. A. Fuller. "Production of murine hybrid-hybridomas secreting bispecific monoclonal antibodies for use in urease-based immunoassays." Clinical Chemistry 34, no. 9 (1988): 1693–96. http://dx.doi.org/10.1093/clinchem/34.9.1890.

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Abstract To produce bispecific antibodies (BiAbs) for enzyme immunoassay (EIA) to replace antibody-enzyme conjugates, we developed a panel of 8-azaguanine/ouabain-resistant anti-urease variant hybridoma cell lines for use in hybridoma-hybridoma fusions. These variants represent mouse immunoglobulin subclasses IgM, IgG1, IgG2a, and IgG2b and have growth rates equal to those of the parental hybridomas. We fused an anti-urease-secreting variant hybridoma with an anti-human choriogonadotropin (hCG)-secreting hybridoma (both of IgG1 subclass) and selected the desired product with growth media conta

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Leibnitz, R. R., P. E. Lipsky, and D. L. Thiele. "Reactivity of hybridomas derived from T cells activated in vivo during graft-versus-host disease." Journal of Immunology 153, no. 11 (1994): 4959–68. http://dx.doi.org/10.4049/jimmunol.153.11.4959.

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Abstract To examine the specificity of T helper cells activated during murine graft-vs-host disease (GVHD), T cell hybridomas from GVHD spleens and livers were generated and analyzed. CTL-depleted C57BL/6 (B6) donor cells were injected into irradiated (B6 x bm12)F1 or (bm1 x bm12)F1 recipient mice. Five or fourteen days later, cells from livers and spleens were fused directly with the TCR-deficient (alpha beta)- BW5147 thymoma line. The in vivo-activated T cells produced hybridomas as efficiently as either T cells activated in a primary mixed lymphocyte reaction or expanded in vitro after isol

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Dzhambazov, Balik, Tsvetelina Batsalova, Patrick Merky, Franziska Lange, and Rikard Holmdahl. "NIH/3T3 Fibroblasts Selectively Activate T Cells Specific for Posttranslationally Modified Collagen Type II." International Journal of Molecular Sciences 24, no. 13 (2023): 10811. http://dx.doi.org/10.3390/ijms241310811.

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It has been shown that synovial fibroblasts (SF) play a key role in the initiation of inflammation and joint destruction, leading to arthritis progression. Fibroblasts may express major histocompatibility complex class II region (MHCII) molecules, and thus, they could be able to process and present antigens to immunocompetent cells. Here we examine whether different types of fibroblasts (synovial, dermal, and thymic murine fibroblasts, destructive LS48 fibroblasts, and noninvasive NIH/3T3 fibroblasts) may be involved in the initiation of rheumatoid arthritis (RA) pathogenesis and can process a

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Sherr, E., D. C. Adelman, A. Saxon, M. Gilly, R. Wall, and N. Sidell. "Retinoic acid induces the differentiation of B cell hybridomas from patients with common variable immunodeficiency." Journal of Experimental Medicine 168, no. 1 (1988): 55–71. http://dx.doi.org/10.1084/jem.168.1.55.

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Human-human B cell hybridomas constructed from B lymphocytes of common variable immunodeficiency (CVI) patients and the nonsecreting cell line WIL2/729 HF consistently secrete low levels of Ig and appear to retain a defect characteristic of the CVI patient's B cells. We assessed the differentiative capacity of retinoic acid (RA) on these hybridomas, as well as on hybridomas constructed from normal B cells and from patients with selective IgA deficiency. RA at concentrations varying between 10(-5) and 10(-9) M augmented IgM secretion 4-20-fold from four of four CVI hybridomas tested, but did no

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Ju, S. T., and M. E. Dorf. "Functional analysis of cloned macrophage hybridomas. IV. Induction and inhibition of mixed lymphocyte responses." Journal of Immunology 134, no. 6 (1985): 3722–30. http://dx.doi.org/10.4049/jimmunol.134.6.3722.

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Abstract A series of macrophage (M phi) hybridomas were generated by fusion of drug-marked P388D1 (H-2d) tumor cells with CKB (H-2k) splenic adherent cells. The ability of this panel of cloned M phi hybridomas expressing various levels of surface Ia antigens to induce allogeneic mixed lymphocytes responses (MLR) was examined. All MLR stimulatory M phi hybridomas expressed surface Ia antigens. However, some Ia+ and all Ia- M phi hybridomas were unable to induce vigorous MLR responses. Furthermore, even after induction of surface Ia antigen expression with Con A supernatants (Con A Sn) or purifi

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van der Schoot, Johan M. S., Felix L. Fennemann, Michael Valente, et al. "Functional diversification of hybridoma-produced antibodies by CRISPR/HDR genomic engineering." Science Advances 5, no. 8 (2019): eaaw1822. http://dx.doi.org/10.1126/sciadv.aaw1822.

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Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete antibodies in the preferred format, species, and isotype. Using this platform, we obtained recombin

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Yang, Y., M. Merćep, C. F. Ware, and J. D. Ashwell. "Fas and activation-induced Fas ligand mediate apoptosis of T cell hybridomas: inhibition of Fas ligand expression by retinoic acid and glucocorticoids." Journal of Experimental Medicine 181, no. 5 (1995): 1673–82. http://dx.doi.org/10.1084/jem.181.5.1673.

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Activation of T cell hybridomas induces a G1/S cell cycle block and apoptosis. We isolated a variant of the 2B4.11 T cell hybridoma that, when activated via the TCR, produced IL-2 and underwent growth inhibition but did not die. Analysis of a variety of cell surface molecules revealed that the variant cell line, termed VD1, expressed very low levels of Fas compared to the wild type cells. Unlike 2B4.11 cells, VD1 cells were not killed by Fas ligand (FasL)-bearing effector cells. To determine if Fas is involved in activation-induced apoptosis, two different reagents that specifically bind Fas w

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Cron, R. Q., A. Ezquerra, J. E. Coligan, B. A. Houlden, J. A. Bluestone, and W. L. Maloy. "Identification of distinct T cell receptor (TCR)-gamma delta heterodimers using an anti-TCR-gamma variable region serum." Journal of Immunology 143, no. 11 (1989): 3769–75. http://dx.doi.org/10.4049/jimmunol.143.11.3769.

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Abstract T cell hybridomas were generated from CD3+, CD4-, CD8- splenocytes and fetal thymocytes. V gamma 1-expressing proteins present on these murine TCR-gamma delta hybridomas were identified by using an anti-TCR V gamma 1 peptide serum. This antiserum specifically immunoprecipitated 41-kDa TCR V gamma-C gamma 4 chains and 31-kDa TCR V gamma-C gamma 1/2 chains from distinct heterodimers expressed on the TCR-gamma delta T cell hybridomas. The RNA from a hybridoma with a 31-kDa TCR-gamma chain hybridized with a V gamma 1 probe but failed to hybridize with a V gamma 2 probe. In contrast, the R

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Vaghela, Arunkumar Ramjibhai, and Tejas H. Ganatra. "A detailed review of immunotherapeutics with a special emphasis on hybridoma technology." American Journal of Biopharmacy and Pharmaceutical Sciences 4 (January 25, 2024): 2. http://dx.doi.org/10.25259/ajbps_13_2023.

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The paper offers a thorough analysis of immunotherapeutics with a focus on hybridomas. It describes how focused and precise treatments for a variety of illnesses, such as cancer, autoimmune disorders, and infectious diseases, have been made possible by immunotherapeutics, which are based on antibody and hybridoma technology. The main therapeutics produced by this method are monoclonal antibodies (mAbs). The article describes the hybridoma technology process, in which a heterogeneous population of cells that produce unique mAbs are created by combining immortalized myeloma cells with B lymphocy

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Greenfield, Edward A. "Hybridoma Screening by Antibody Capture: Flow Cytometry/FACS with Permeabilized Cells to Detect Intracellular Binding." Cold Spring Harbor Protocols 2022, no. 6 (2022): pdb.prot103085. http://dx.doi.org/10.1101/pdb.prot103085.

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Flow cytometry or fluorescence-activated cell sorting (FACS) can be used to identify hybridomas secreting monoclonal antibodies to internal cellular proteins, but the cells must be permeabilized before the hybridoma supernatants are applied. In using this technique, useful controls are positive and negative cell lines with primary and secondary antibodies as well as positive and negative cell lines with secondary antibody alone.

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Kanagawa, O., and R. Maki. "Inhibition of MHC class II-restricted T cell response by Lyt-2 alloantigen." Journal of Experimental Medicine 170, no. 3 (1989): 901–12. http://dx.doi.org/10.1084/jem.170.3.901.

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T cell hybridomas were established by fusing a CD8+ V beta 8.1+ CTL clone and a CD4+ V beta 8.1+ helper T lymphocyte (HTL) clone to the thymoma cell line BW5147. In contrast to the HTL x BW hybridomas, which retain the same antigen specificity as the original T cell clone, the CTL x BW hybridomas lost the class I MHC-restricted antigen response but acquired a new specificity to Mlsa antigen. Mlsa reactivity of CTL x BW hybridomas was shown to be mediated by the CTL TCR as assayed by inhibition using an anticlonotypic antibody to the CTL clone. Since hybridomas established with BW5147 lose CD8

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Pornnoppadol, Ghasidit, Boya Zhang, Alec A. Desai, et al. "A hybridoma-derived monoclonal antibody with high homology to the aberrant myeloma light chain." PLOS ONE 16, no. 10 (2021): e0252558. http://dx.doi.org/10.1371/journal.pone.0252558.

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The identification of antibody variable regions in the heavy (VH) and light (VL) chains from hybridomas is necessary for the production of recombinant, sequence-defined monoclonal antibodies (mAbs) and antibody derivatives. This process has received renewed attention in light of recent reports of hybridomas having unintended specificities due to the production of non-antigen specific heavy and/or light chains for the intended antigen. Here we report a surprising finding and potential pitfall in variable domain sequencing of an anti-human CD63 hybridoma. We amplified multiple VL genes from the

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Hagiwara, H., T. Yokota, J. Luh, et al. "The AKR thymoma BW5147 is able to produce lymphokines when stimulated with calcium ionophore and phorbol ester." Journal of Immunology 140, no. 5 (1988): 1561–65. http://dx.doi.org/10.4049/jimmunol.140.5.1561.

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Abstract We produced the T cell hybridoma D9C1.12.17 by fusing an IL-4-producing T cell clone D9.1Hi with the AKR thymoma BW5147. The resulting hybridoma produced IL-2 as well as IL-4 even though none of the parental cells produced IL-2 after stimulation with Con A. The production of IL-2 was confirmed at the mRNA level by using an S1 nuclease protection assay. Further analysis indicated that Con A-induced IL-2 production was a common phenomenon among T cell hybridomas derived from this fusion. Although BW5147 does not produce detectable lymphokines after Con A stimulation, this line was able

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Barka, T., E. W. Gresik, and H. van der Noen. "Monoclonal antibodies against rat saliva and salivary gland antigens." Journal of Histochemistry & Cytochemistry 33, no. 3 (1985): 209–18. http://dx.doi.org/10.1177/33.3.2579121.

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Hybridomas were produced by the fusion of NS1 myeloma cells with spleen cells of a BALB/c mouse immunized with rat submandibular saliva. Growth of hybridomas was evident in 60/96 wells, and colonies secreting antibodies against saliva components were identified in 20 wells by using a solid phase enzyme-linked immunoassay. Cloning of cells from 12 wells yielded originally 43 hybridoma cell lines secreting anti-saliva antibodies. After recloning, one hybridoma (4Cl3) was selected for further studies. The hybridoma (4Cl3) cells were grown as ascites tumors, and the antibodies were purified from t

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Kuchroo, V. K., M. Minami, B. Diamond, and M. E. Dorf. "Functional analysis of cloned macrophage hybridomas. VI. Differential ability to induce immunity or suppression." Journal of Immunology 141, no. 1 (1988): 10–16. http://dx.doi.org/10.4049/jimmunol.141.1.10.

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Abstract We previously screened a series of macrophage hybridomas derived from fusion of P388D1 (H-2d) tumor cells with CKB (H-2k) splenic adherent cells for their ability to induce I-J restricted Ts cell responses. One Ia+ macrophage clone (63) consistently induced Ag-specific, I-J-restricted Ts. To evaluate whether macrophage hybridoma 63 also induced delayed-type hypersensitivity (DTH) immunity, mice were immunized with hapten-coupled macrophage hybridoma cells. Hapten-coupled splenic adherent cells and control macrophage hybridomas induced significant primary DTH responses, whereas hapten-

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Gu, J. J., J. V. Harriss, K. Ozato, and P. D. Gottlieb. "Induction by concanavalin A of specific mRNAs and cytolytic function in a CD8-positive T cell hybridoma." Journal of Immunology 153, no. 10 (1994): 4408–17. http://dx.doi.org/10.4049/jimmunol.153.10.4408.

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Abstract A previous report from this laboratory described the production of CD8+, class I-specific T cell hybridomas which developed specific cytolytic activity and the ability to secrete IL-2 upon Con A or specific Ag stimulation. Unlike normal lymphocytes or long-term CTL lines for which exposure to Ag triggers both differentiation and proliferation, T cell hybridoma lines can be activated functionally against a background of continuous proliferation. They therefore provide a unique system with which to study the molecular events involved in the induction of cytolytic function. The expressio

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Heilmann, Katja, and Pamela Holzlöhner. "Generation of antigen-specific monoclonal antibodies by in vitro immunization (TECH2P.904)." Journal of Immunology 194, no. 1_Supplement (2015): 206.14. http://dx.doi.org/10.4049/jimmunol.194.supp.206.14.

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Abstract Monoclonal antibodies are one of the most important tools in biomedicine but the generation by hybridoma technology is very time consuming and laborious. The adaptation of the process to an in vitro approach is therefore a smart alternative to reduce the disadvantages of hybridoma technology. In this study a protocol for in vitro immunization was developed which enables hematopoietic stem cells to differentiate into naïve dendritic cells. These cells were specifically activated with the antigen of interest and cocultivated with naïve B and T lymphocytes in order to generate antigen sp

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Makoul, G. T., D. R. Robinson, A. K. Bhalla, and L. H. Glimcher. "Prostaglandin E2 inhibits the activation of cloned T cell hybridomas." Journal of Immunology 134, no. 4 (1985): 2645–50. http://dx.doi.org/10.4049/jimmunol.134.4.2645.

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Abstract To minimize complicating interactions inherent in heterogeneous cell populations, we used a panel of cloned murine autoreactive (E8.A1) and antigen-specific (HEL.C10, HEL.B14) T cell hybridomas to examine the effect of prostaglandin E2 (PGE2) on T cell activation. These T cells secrete interleukin 2 (IL 2) when co-cultured with a cloned population of I region-matched stimulator cells (TA3), or with mitogenic signals in the absence of TA3 stimulator cells. Physiologic concentrations of PGE2 inhibited the induction of IL 2 secretion by the T cell hybridomas tested, when they were activa

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Minami, M., H. Nariuchi, H. Kawasaki, and M. E. Dorf. "The role of class II MHC molecules in the activation of class I-reactive T cell hybridomas." Journal of Immunology 136, no. 9 (1986): 3341–45. http://dx.doi.org/10.4049/jimmunol.136.9.3341.

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Abstract To examine the role of Ia molecules in T cell responses to allo-class I major histocompatibility antigens, a series of allo-class I-reactive T cell hybridomas was established. Of 134 T cell hybridomas obtained from the fusion of C3H/HeJm or B10.HTT T cells stimulated with C57BL/6 splenocytes, nine T cell hybridomas were reactive to class I antigens and 126 T cell hybridomas were reactive to class II antigens. Six of the nine IL 2-producing T cell hybridomas were further analyzed: five mapped to H-2Kb and the other mapped to H-2Db. Three of these T cell hybridomas, HTB-157.7, HTB-176.1

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Jaffredo, T., A. F. Horwitz, C. A. Buck, P. M. Rong, and F. Dieterlen-Lievre. "Myoblast migration specifically inhibited in the chick embryo by grafted CSAT hybridoma cells secreting an anti-integrin antibody." Development 103, no. 3 (1988): 431–46. http://dx.doi.org/10.1242/dev.103.3.431.

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We report a teratological method in which mouse hybridoma cells are grafted into a chick host. CSAT (Cell Substratum ATtachment) hybridoma was used. It produces an antibody directed against the avian integrin complex. The grafts were performed during the second and third days of incubation either at the level of the somites or in the coelom of the chick embryo. The anomalies were revealed by means of a monoclonal antibody that recognizes myogenic cells as soon as they become committed in the myotome. When embryos were grafted at the level of the somites, body wall muscles failed to develop on

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Raychaudhuri, S., Y. Saeki, H. Fuji, and H. Kohler. "Tumor-specific idiotype vaccines. I. Generation and characterization of internal image tumor antigen." Journal of Immunology 137, no. 5 (1986): 1743–49. http://dx.doi.org/10.4049/jimmunol.137.5.1743.

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Abstract The concept of idiotype vaccines against tumor-associated antigens (TAA) was tested in the DBA/2 L1210 lymphoma subline, L1210/GZL. Monoclonal antibodies against a TAA that cross-reacts with the envelope glycoprotein gp52 of the mammary tumor virus were used to make hybridoma anti-idiotype antibodies (Ab2). In this report we describe the characterization of monoclonal anti-idiotypic antibodies against the combining site of 11C1 (Ab1), which recognizes a shared determinant of gp52 of mouse mammary tumor virus (MMTV) and the TAA of L1210/GZL. Hybridomas expressing the internal image of

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Muralidhar, G., H. Sepulveda, A. J. Feeney, R. Riblet, and R. W. Dutton. "Restricted IgH V gene usage in the response to the ese epitope on "Hi" sheep red blood cells." Journal of Immunology 149, no. 11 (1992): 3574–79. http://dx.doi.org/10.4049/jimmunol.149.11.3574.

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Abstract We had previously shown that the in vitro antibody response to a single epitope (ese; extra sheep E Ag) present on some sheep E but absent from others could be monitored by assay of the plaque-forming cell response on both Lo3 and Hi SRBC. We had shown also that the response was seen only in certain strains of mice and that the gene(s) controlling the response mapped to the IgH V region of the IgH chain complex. An additional feature of the response is that it is only seen in vitro and is absent and, we hypothesize, is suppressed in vivo. The strain distribution of the response to the

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Maillère, B., J. Cotton, G. Mourier, M. Léonetti, S. Leroy, and A. Ménez. "Role of thiols in the presentation of a snake toxin to murine T cells." Journal of Immunology 150, no. 12 (1993): 5270–80. http://dx.doi.org/10.4049/jimmunol.150.12.5270.

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Abstract We isolated and characterized two T hybridomas specific for a highly stable snake toxic protein. One hybridoma, called T1C9, is I-E(d)-restricted and stimulated by both the native and reduced and carboxymethylated (RCM) toxins and by synthetic fragments containing the region 24-36. The other hybridoma, called T1B2, is I-A(d)-restricted and stimulated by the native toxin, only. Neither the RCM toxin nor any of the initial synthetic peptides used in our study could stimulate it. We show that this lack of effect is associated with the presence, in the epitope-containing fragment, of irre

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Martel, F., R. Bazin, S. Verrette, and R. Lemieux. "Characterization of higher avidity monoclonal antibodies produced by murine B-cell hybridoma variants selected for increased antigen binding of membrane Ig." Journal of Immunology 141, no. 5 (1988): 1624–29. http://dx.doi.org/10.4049/jimmunol.141.5.1624.

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Abstract Somatic mutation in the Ig genes plays a major role in the increase of antibody affinity observed in secondary immunologic responses. It has been shown that the mechanism responsible for the high rate of somatic mutation in the Ig genes was active not only in normal B lymphocytes but also in B-cell hybridomas secreting mAb. Also, it has been reported that B-cell hybridomas were positive for membrane Ig of the same specificity as the secreted mAb. The presence of membrane Ig suggested that somatic variants secreting mAb of higher affinity could be selected by the increased capacity of

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Goldfien, R. D., P. J. Chen, T. J. Kipps, et al. "Genetic analysis of human B cell hybridomas expressing a cross-reactive idiotype." Journal of Immunology 138, no. 3 (1987): 940–44. http://dx.doi.org/10.4049/jimmunol.138.3.940.

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Abstract We have examined the genetic basis for the expression of a human cross-reactive idiotype (CRI) commonly found on monoclonal IgM rheumatoid factors. The CRI was identified with a monoclonal antibody (17.109) and has been localized previously to the kappa-variable region. By using the human lymphoblastoid cell line WI-L2-729-HF2, and mononuclear cells from several sources, a panel of hybridomas was generated that produced 17.109 CRI-positive Ig. A recently cloned human germ-line V kappa III gene, Humkv305, served as a probe to identify genes which were rearranged and expressed in 17.109

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Glickstein, L., S. Macphail, and O. Stutman. "Uncoupling IL-2 production from apoptosis and TNF production by changing the signal through the TCR." Journal of Immunology 156, no. 6 (1996): 2062–67. http://dx.doi.org/10.4049/jimmunol.156.6.2062.

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Abstract T cells may discriminate between stimuli in a variety of ways, including the presence of cytokines or other costimulatory signals, the type of Ag (peptide, superantigen, or allorecognition), or the magnitude of the signal through the TCR. We have used anti-CD3 stimulation of T hybridomas to examine signals generated through the TCR in the absence of exogenous APCs. Soluble whole anti-CD3, but not F(ab')2 anti-CD3, was able to stimulate the T hybridomas to produce IL-2. Plastic-bound anti-CD3, in contrast, stimulated TNF production, G1 arrest, and apoptosis by the T hybridoma. Engageme

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Teng, Man, Jin-Ling Liu, Qin Luo, et al. "Efficient Cross-Screening and Characterization of Monoclonal Antibodies against Marek’s Disease Specific Meq Oncoprotein Using CRISPR/Cas9-Gene-Edited Viruses." Viruses 15, no. 4 (2023): 817. http://dx.doi.org/10.3390/v15040817.

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Marek’s disease (MD) caused by pathogenic Marek’s disease virus type 1 (MDV−1) is one of the most important neoplastic diseases of poultry. MDV−1-encoded unique Meq protein is the major oncoprotein and the availability of Meq-specific monoclonal antibodies (mAbs) is crucial for revealing MDV pathogenesis/oncogenesis. Using synthesized polypeptides from conserved hydrophilic regions of the Meq protein as immunogens, together with hybridoma technology and primary screening by cross immunofluorescence assay (IFA) on Meq-deleted MDV−1 viruses generated by CRISPR/Cas9-gene editing, a total of five

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