Characterization of the primary IgM response to GAT and GT

Conditions required for the detection of IgM antibodies

C. Waltenbaugh, A. Dessein, B. Benacerraf

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Primary IgM antibody responses to synthetic linear copolymers of L-glutamic acid, L-tyrosine, and L-alanine were investigated. The appearance of primary IgM anti-GAT antibodies was detected in BALB/c mice by using a solid phase of radioimmunoassay (SPRIA) procedure. This finding was verified for GAT in responder mice and GAT-MBSA and GT-MBSA in nonresponder mice in an indirect plaque forming cell (PFC) assay by using a rabbit antiserum directed against the μλ myeloma protein, MOPC 104E. Facilitated IgM PFC could be inhibited by a purified μκ myeloma protein, TEPC 183. Maximal facilitated IgM plaque response was found to precede the IgG response by several days. A direct plaque assay was developed for the detection of IgM anti-GAT plaques using poly-L-lysine (PLL) to couple GAT to sheep erythrocytes (SRBC). GAT-SRBC coupled by the PLL method optimally couple 4 to 5 times less antigen to the indicator cell surface than does the CrCl3-coupling method routinely employed in our laboratory. These findings were extended to a conventional antigen, chicken gamma globulin (CγG). We found that a less dense epitope coat on the indicator cell surface favors detection of direct IgM PFC, whereas a more densely coated indicator cell favors the detection of facilitated IgM and IgG PFC responses.

Original languageEnglish (US)
Pages (from-to)27-33
Number of pages7
JournalJournal of Immunology
Volume122
Issue number1
StatePublished - Jan 1 1979

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Immunoglobulin M
Antibodies
Myeloma Proteins
Lysine
Immunoglobulin G
Antigens
gamma-Globulins
GAT
Alanine
Antibody Formation
Radioimmunoassay
Tyrosine
Immune Sera
Epitopes
Glutamic Acid
Anti-Idiotypic Antibodies
Chickens
Sheep
Erythrocytes
Rabbits

ASJC Scopus subject areas

  • Immunology

Cite this

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abstract = "Primary IgM antibody responses to synthetic linear copolymers of L-glutamic acid, L-tyrosine, and L-alanine were investigated. The appearance of primary IgM anti-GAT antibodies was detected in BALB/c mice by using a solid phase of radioimmunoassay (SPRIA) procedure. This finding was verified for GAT in responder mice and GAT-MBSA and GT-MBSA in nonresponder mice in an indirect plaque forming cell (PFC) assay by using a rabbit antiserum directed against the μλ myeloma protein, MOPC 104E. Facilitated IgM PFC could be inhibited by a purified μκ myeloma protein, TEPC 183. Maximal facilitated IgM plaque response was found to precede the IgG response by several days. A direct plaque assay was developed for the detection of IgM anti-GAT plaques using poly-L-lysine (PLL) to couple GAT to sheep erythrocytes (SRBC). GAT-SRBC coupled by the PLL method optimally couple 4 to 5 times less antigen to the indicator cell surface than does the CrCl3-coupling method routinely employed in our laboratory. These findings were extended to a conventional antigen, chicken gamma globulin (CγG). We found that a less dense epitope coat on the indicator cell surface favors detection of direct IgM PFC, whereas a more densely coated indicator cell favors the detection of facilitated IgM and IgG PFC responses.",
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Characterization of the primary IgM response to GAT and GT : Conditions required for the detection of IgM antibodies. / Waltenbaugh, C.; Dessein, A.; Benacerraf, B.

In: Journal of Immunology, Vol. 122, No. 1, 01.01.1979, p. 27-33.

Research output: Contribution to journalArticle

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AB - Primary IgM antibody responses to synthetic linear copolymers of L-glutamic acid, L-tyrosine, and L-alanine were investigated. The appearance of primary IgM anti-GAT antibodies was detected in BALB/c mice by using a solid phase of radioimmunoassay (SPRIA) procedure. This finding was verified for GAT in responder mice and GAT-MBSA and GT-MBSA in nonresponder mice in an indirect plaque forming cell (PFC) assay by using a rabbit antiserum directed against the μλ myeloma protein, MOPC 104E. Facilitated IgM PFC could be inhibited by a purified μκ myeloma protein, TEPC 183. Maximal facilitated IgM plaque response was found to precede the IgG response by several days. A direct plaque assay was developed for the detection of IgM anti-GAT plaques using poly-L-lysine (PLL) to couple GAT to sheep erythrocytes (SRBC). GAT-SRBC coupled by the PLL method optimally couple 4 to 5 times less antigen to the indicator cell surface than does the CrCl3-coupling method routinely employed in our laboratory. These findings were extended to a conventional antigen, chicken gamma globulin (CγG). We found that a less dense epitope coat on the indicator cell surface favors detection of direct IgM PFC, whereas a more densely coated indicator cell favors the detection of facilitated IgM and IgG PFC responses.

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