Efficient production of IGG human monoclonal antibodies by lymphocytes stimulated by lipopolysaccharide, pokeweed mitogen, and interleukin 4

Yoshinari, Kaoru; Arai, Kenji; Kimura, Hideki; Matsumoto, Kunio; Yamaguchi, Yutaka

doi:10.1007/bf02722964

Cited by 4 publications

(4 citation statements)

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“…Table 2 shows the results on the cell proliferation and Ab (Ig) production, after 2-3 weeks of incubation of the hybridomas, which had been obtained by electrofusion of mouse myeloma cells P3X63Ag8.653 and the lymphocytes previously incubated for 5 d with either LPS or SACI. In the case of no addition of mitogens, hybridomas were hardly generated as reported previously (Yoshinari et al, 1996).…”

Section: Lymphocyte Subsetssupporting

confidence: 64%

“…Regional lymph nodes from patients with primary lung cancer were obtained asceptically during surgery as described previously (Yoshinari et al, 1995;Yoshinari et al, 1996). A single-cell suspension (lymphocytes) in 10% FCS-supplemented RPMI 1640 (Flow Laboratories) medium was prepared by passing the lymph nodes through sterile steel mesh (#200) after teasing with scissors.…”

Section: Lymphocyte Preparationmentioning

confidence: 99%

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et al. 1999

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Regional lymph node lymphocytes from five patients with primary lung cancer were analyzed for subset composition, and exposed in vitro to the polyclonal human B cell mitogen Staphylococcus aureus Cowan I (SACI) or the murine B cell mitogen lipopolysaccharide (LPS) and then fused with mouse myeloma cells for investigation at the clonal level of their antibody (Ab) production and its statistical relation to the original subset composition. No correlation was found between the proportion of CD19+, CD23+, or CD3+ cells in the lymphocyte sample prior to its exposure to either SACI or LPS, and the Ab production efficiency, defined as the ratio of the number of Ab producing wells to the total number of proliferating wells. For lymphocytes exposed to LPS, however, a strong correlation (r = 0.931, p = 0.02) was observed between the Ab production efficiency and the ratio of CD8+ to CD3+ cells (CD8/CD3) in the original sample at least within the ranges studied (CD8/CD3 = 0.216-0.288). For those exposed to SACI, no correlation was found between the Ab production efficiency and the CD8/CD3 ratio (r = 0.881, p = 0.12) or the proportion of CD8+ cells (r = 0.808, p = 0.19) in the original sample. These results suggest that the repertoire of B cells responsive to LPS is different at least in part from the repertoire responsive to SACI and that the ratio CD8/CD3 could serve as a practical predictor for Ab production by human lymphocytes stimulated with LPS.

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Section: Lymphocyte Subsetssupporting

confidence: 64%

Section: Lymphocyte Preparationmentioning

confidence: 99%

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et al. 1999

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“…The results of the present study indicate that LPS is effective for such activation, at least when used in combination with lymphokines such as IL-4, IL-6 and IL-7. On this basis, we have now begun a systematic study to determine the combinations of mitogens and lymphokines most effective for this purpose (Yoshinari et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

Generation of human monoclonal antibodies recognising membranous antigens of the lung adenocarcinoma cell line A549 using an AMeX immunohistostaining method

Yoshinari

Kimura²,

Arai³

et al. 1996

Br J Cancer

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Summary Four monoclonal antibodies (MAbs) from hybridoma obtained by in vitro stimulation of regional lymph node lymphocytes from lung cancer patients and electrofusion of the stimulated cells with murine or human -mouse myeloma cells were reactive to lung cancer cells in enzyme-linked immunoabsorbent assay, and to lung cancer tissue in immunohistochemical analysis using acetone-methyl benzoate-xylene (AMeX) fixed tissue and in immunofluorescence analysis. Three of the MAbs (designated ZLG40, 27D57 and 28K29) recognised cell-surface antigens of the lung adenocarcinoma cell line A549 and the remaining one (designated 29D38) recognised nuclear membrane antigens of the same cell line. The three surface-binding MAbs showed a significant complement-dependent cytotoxicity (CDC) to the A549 cells, but the membrane-binding 29D38 showed no CDC to the A549 cells. Western blotting of the extracts of the A549 or PC6 (small-cell lung cancer) cell lines by the four MAbs showed a 28K29 antigen band at M, of approximately 600 000 (± 2-ME), a ZLG40 antigen band at Mr 50 000 (±2-ME), and one 29D38 antigen band at Mr of more than 1 000 000 (-2-ME) and Mr between 20 000 and 80 000 (+ 2-ME), but no detectable band for 27D57 antigen.

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“…Fusion is then induced by one or more bursts of direct current. Electrofusion has been used with a variety of fusion partner cell lines, including lymphoblastoid, heteromyeloma and murine myeloma cell lines (Pratt et al 1987;Foung et al 1990;Yoshinari et al 1996). Three studies have directly compared the efficiency of electrofusion and polyethylene glycol (PEG) fusion, estimating an apparent superiority of electrofusion of 4-to 100-fold, with a maximal calculated fusion rate of approximately 1 cell per 1,000 input human B cells (Perkins et al 1991;Krenn et al 1995;Panova and Gustafsson 1995).…”

Section: Electrofusion and Hybrid Cell Culturementioning

confidence: 99%

Exploring the Native Human Antibody Repertoire to Create Antiviral Therapeutics

Dessain

Adekar

Berry³

2008

Current Topics in Microbiology and Immunology

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Native human antibodies are defined as those that arise naturally as the result of the functioning of an intact human immune system. The utility of native antibodies for the treatment of human viral diseases has been established through experience with hyperimmune human globulins. Native antibodies, as a class, differ in some respects from those obtained by recombinant library methods (phage or transgenic mouse) and possess distinct properties that may make them ideal therapeutics

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Efficient production of IGG human monoclonal antibodies by lymphocytes stimulated by lipopolysaccharide, pokeweed mitogen, and interleukin 4

Cited by 4 publications

References 29 publications

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Generation of human monoclonal antibodies recognising membranous antigens of the lung adenocarcinoma cell line A549 using an AMeX immunohistostaining method

Exploring the Native Human Antibody Repertoire to Create Antiviral Therapeutics

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