[5] Method for generating a high frequency of hybridomas producing monoclonal IgA antibodies

Colwell, Dawn E.; Michalek, Suzanne M.; McGhee, Jerry R.

doi:10.1016/0076-6879(86)21007-1

Cited by 11 publications

(3 citation statements)

References 19 publications

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“…Our results establish that PP cells are effective in generating IgA-secreting hybridomas when directly fused with myeloma cells. IgA-producing hybridomas have been generated previously using spleen cells from mucosally immunized animals (8,18), but the technique is not always successful (9). Cells from MLN and thoracic duct lymph (31,41) or mixed cells from MLN and PPs (13) have been used successfully, and it was suggested that PP lymphocytes alone may be effective in generating IgA-secreting hybridomas (41).…”

Section: Pp Cells From Orally Immunized Mice As a Source Of Polymeric Iga Antibodiesmentioning

confidence: 99%

Binding and transepithelial transport of immunoglobulins by intestinal M cells: demonstration using monoclonal IgA antibodies against enteric viral proteins.

Weltzin

Lucia-Jandris

Michetti

et al. 1989

The Journal of Cell Biology

228

126

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Abstract. M cells of intestinal epithelia overlying lymphoid follicles endocytose luminal macromolecules and microorganisms and deliver them to underlying lymphoid tissue. The effect of luminal secretory IgA antibodies on adherence and transepithelial transport of antigens and microorganisms by M cells is unknown. We have studied the interaction of monoclonal IgA antibodies directed against specific enteric viruses, or the hapten trinitrophenyl (TNP), with M cells. To produce monospecific IgA antibodies against mouse mammary tumor virus (MMTV) and reovirus type 1, Peyer's patch cells from mucosally immunized mice were fused with myeloma cells, generating hybridomas that secreted virus-specific IgA antibodies in monomeric and polymeric forms. One of two anti-MMTV IgA antibodies specifically bound the viral surface glycoprotein gp52, and 3 of 10 antireovirus IgA antibodies immunoprecipitated sigma 3 and mu lc surface proteins. 35S-labeled IgA antibodies injected intravenously into rats were recovered in bile as higher toolecular weight species, suggesting that secretory component had been added on passage through the liver. Radiolabeled or colloidal gold-conjugated mouse IgA was injected into mouse, rat, and rabbit intestinal loops containing Peyer's patches. Light microscopic autoradiography and EM showed that all IgA antibodies (antivirus or anti-TNP) bound to M cell luminal membranes and were transported in vesicles across M cells. IgA-gold binding was inhibited by excess unlabeled IgA, indicating that binding was specific. IgGgold also adhered to M cells and excess unlabeled IgG inhibited IgA-gold binding; thus binding was not isotype-specific. Immune complexes consisting of monoclonal anti-TNP IgA and TNP-ferritin adhered selectively to M cell membranes, while TNP-ferritin alone did not. These results suggest that selective adherence of luminal antibody to M cells may facilitate delivery of virus-antibody complexes to mucosal l~m-phoid tissue, enhancing subsequent secretory immune responses or facilitating viral invasion.

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Section: Pp Cells From Orally Immunized Mice As a Source Of Polymeric Iga Antibodiesmentioning

confidence: 99%

Binding and transepithelial transport of immunoglobulins by intestinal M cells: demonstration using monoclonal IgA antibodies against enteric viral proteins.

Weltzin

Lucia-Jandris

Michetti

et al. 1989

The Journal of Cell Biology

228

126

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“…The review "Refinement of Monoclonal Antibody Production and Animal Well-being" by McGuill and Rowan (ILAR News 31[l]:7-10, 1989) correctly demonstrates that significant progress is being made but that important elements are still lacking. Especially encouraging are the recent papers by Colwell et al (1986), Hoogenraad and Wraight (1986), and Kwan et al (1980), which show that in ascites production, smaller volumes of pristane used in priming are as effective as standard volumes. The review authors also point out an important missing element: studies to date have not determined dosages of priming agents that produce high titered ascitic fluid yields with minimal stress to the animal.…”

Section: Refinement Of Mab Production a Welcome Trendmentioning

confidence: 93%

Refinement of MAB Production a Welcome Trend

Amyx¹

1989

ILAR Journal

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“…One study demonstrated no significant difference in ascites volumes in animals receiving 0. 1 vs. 0.5 ml pristane (Hoogenraad and Wraight, 1986), and researchers have reported success using 0.2 ml pristane (Kwan et al, 1980;Colwell et al, 1986). Other authors suggest that smaller volumes of pristane may reduce the potential for distress associated with its irritant properties (Amyx, 1987;McGuill and Rowan, 1989).…”

Section: Priming Agentsmentioning

confidence: 99%