Generation of human monoclonal antibodies to cancer-associated antigens using limited numbers of patient lymphocytes

Smith, Lloyd H.; Yin, Amy X.; Bieber, Marcia M.; Teng, Nelson N.H.

doi:10.1016/0022-1759(87)90274-2

Cited by 9 publications

(4 citation statements)

References 16 publications

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“…Other investigators report levels of 3-6 ug/ml from human-human hybridomas (4,36) and up to 20 pg/ml from human-murine and human-heteromyeloma hybrids (35,42).…”

Section: Discussionmentioning

confidence: 95%

“…Although uncloned lymphoblastoid cells have previously been used to generate hybridomas (35), the two types of cell lines have not been directly compared. Stable lymphoblastoid cell lines are only rarely established.…”

Section: Discussionmentioning

confidence: 99%

“…At day ten and every five days thereafter, 0.5 ml media was removed from the wells and replaced with fresh RPMI 1640 containing 10% FBS. Most fusions had hybrids evident by 21 days and were screened by [28][29][30][31][32][33][34][35] days. Hybrid cells secreting T antigen-specific immunoglobulin were cloned immediately after screening at three and ten cells per well with irradiated murine spleen cells (2xl05/well).…”

Section: Fusionmentioning

confidence: 99%

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Stability of Specific Immunoglobulin Secretion by EBV-Transformed Lymphoblastoid Cells and Human-Murine Heterohybridomas

Glasky

Reading²

1989

Hybridoma

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We have examined variables leading to the generation of stable, antigen-specific, human immunoglobulin-secreting cell lines. Peripheral blood B lymphocytes enriched for Thomsen-Friedenreich antigen (T antigen)-specific cells were transformed with Epstein-Barr virus. Lymphoblastoid cells (LC) reactive with T antigen were either expanded without cloning or cloned at limiting dilution and then fused with murine 653 cells. Uncloned LCs from three transformations secreting polyclonal anti-T antibody (7-18 micrograms/ml/10(6) cells/24 hr total immunoglobulin) were subcultured at 100 cells/well, and T antigen-reactive cultures pooled. These cultures quickly lost specific antibody secretion, presumably due to overgrowth by clones of unknown specificity. T antigen-reactive LCs that were cloned three times at limiting dilution secreted 0.2 - 6.1 micrograms/ml/10(6) cells/24 hr but died or stopped secreting specific immunoglobulin after 77 to 155 days in culture. Pooling T antigen-reactive clones after each cloning step did not increase the long term stability compared to unpooled clones (p = 0.2). Fusions between cloned LCs and 653 cells failed to yield viable hybrids in nine of ten attempts with seven different LC lines. In contrast, fusion of uncloned LCs and 653 cells resulted in the generation of viable immunoglobulin-secreting heterohybrids in 22 of 24 fusions. The heterohybridomas produced from fusion of uncloned T antigen-reactive cultures with 653 cells secreted significantly more antibody (frequency of cell lines secreting greater than 2 micrograms/ml/10(6) cells/24 hr, p less than 0.01) and higher titers of antibody (frequency of cell lines secreting greater than four hemagglutination units of T antigen-specific antibody, p less than 0.03) than cloned lymphoblastoid cells. The hybrids maintained specific immunoglobulin secretion for longer in culture than either cloned or uncloned lymphoblastoid cell lines (p less than 0.001).

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“…Other investigators report levels of 3-6 ug/ml from human-human hybridomas (4,36) and up to 20 pg/ml from human-murine and human-heteromyeloma hybrids (35,42).…”

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Fusionmentioning

confidence: 99%

See 1 more Smart Citation

Stability of Specific Immunoglobulin Secretion by EBV-Transformed Lymphoblastoid Cells and Human-Murine Heterohybridomas

Glasky

Reading²

1989

Hybridoma

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“…Serum concentrations of IgA and IgM levels were significantly higher in patients than in controls. Generation of Igs have been implicated in contributing to the disease course of multiple cancers, 36 and may serve as useful biomarker of disease activity and/or prognostic. Our results corroborate those of the previous study 37 regarding the significant increase in IgA as well as IgM levels.…”

Section: Discussionmentioning

confidence: 99%

Clinical association of baseline levels of conjugated dienes in low-density lipoprotein and nitric oxide with aggressive B-cell non-Hodgkin lymphoma and their relationship with immunoglobulins and Th1-to-Th2 ratio

Haddouche

Warda

Hadjidj

et al. 2016

JBM

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ObjectiveThe aim of this study was to highlight the clinical association of baseline levels of conjugated dienes in low-density lipoprotein (LDL-BCD) and nitric oxide (NO) with immunoglobulins (Igs) and T helper (Th)1/Th2 ratio in patients with newly diagnosed B-cell non-Hodgkin lymphoma (NHL).Patients and methodsThirty-two newly diagnosed patients with aggressive B-cell NHL and 25 age-, sex-, and body-mass-index-matched healthy controls were randomly selected for a cross-sectional case–control study conducted at the Hematology Department of Tlemcen Medical Centre University (northwest of Algeria).ResultsCirculating levels of LDL-BCD and NO and those of IgA and IgM were significantly higher in patients than in controls. The levels of Th1/Th2 ratio and plasma total antioxidant capacity were significantly lower in patients compared with controls, while malondialdehyde and protein carbonyl levels were significantly higher in patients. B-cell NHL was significantly associated with high levels of LDL-BCD from 25th to 75th percentile (25th percentile: relative risk [RR] =2.26, 95% confidence interval [CI] 1.42–3.59, P=0.014; 50th percentile: RR =2.84, 95% CI 1.72–4.68, P<0.001; 75th percentile: RR =5.43, 95% CI 2.58–11.42, P<0.001). Similarly, the disease was significantly associated with high levels of NO production from 25th to 75th percentile (25th percentile: RR =2.07, 95% CI 1.25–3.44, P=0.024; 50th percentile: RR =2.78, 95% CI 1.63–4.72, P<0.001; 75th percentile: RR =4.68, 95% CI 2.21–9.91, P<0.001). Moreover, LDL-BCD levels were positively and significantly correlated with interferon (IFN)-γ, whereas NO levels were inversely and significantly correlated with IFN-γ and Th1/Th2 ratio.ConclusionLDL-BCD and NO production seem to be associated with aggressive B-cell NHL and alteration of Th1/Th2 ratio. Our results have to be examined using ex vivo mechanistic studies leading to further investigations of these parameters, with an interest in the link between Epstein–Barr virus infection and NO and immunoglobulins.

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Enzyme-linked immunosorbant assay in the rheumatological laboratory

Charles

Maini

1996

Manual of Biological Markers of Disease

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Generation of human monoclonal antibodies to cancer-associated antigens using limited numbers of patient lymphocytes

Cited by 9 publications

References 16 publications

Stability of Specific Immunoglobulin Secretion by EBV-Transformed Lymphoblastoid Cells and Human-Murine Heterohybridomas

Stability of Specific Immunoglobulin Secretion by EBV-Transformed Lymphoblastoid Cells and Human-Murine Heterohybridomas

Clinical association of baseline levels of conjugated dienes in low-density lipoprotein and nitric oxide with aggressive B-cell non-Hodgkin lymphoma and their relationship with immunoglobulins and Th1-to-Th2 ratio

Enzyme-linked immunosorbant assay in the rheumatological laboratory

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