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The influence of an X chromosome-linked immune deficiency gene (xid) on several properties of the anti-azophenylarsonate (Ar) antibody responses of (CBA/N x A/J)F1 (NAF1) mice was examined. With respect to response magnitude, it was found that male, xid-expressing NAF1 mice showed about 1/3 the concentration of serum anti-Ar antibody as normal female NAF1 mice in hyperimmune responses to Brucella abortus (BA)-Ar. In responses induced by keyhole limpet hemocyanin (KLH)-Ar, males showed responses of about 1/8 to 1/2 the female levels, depending on the assay time point. The kinetics of the latter response were identical in mice of the two sexes. No significant difference could be detected in the time-dependent avidity maturation of the anti-Ar antibody elicited by KLH-Ar in xid vs. normal mice. The isotype profile of the day 38 anti-Ar primary response elicited by KLH-Ar in male NAF1 mice differed from that of the female mice in two ways: IgG2a levels were depressed, and a significantly lower number of the male mice demonstrated detectable IgG3 anti-Ar antibody production. The primary focus of htis work was to determine the effect of xid on the expression of the major cross-reactive idiotype--CRIA--of A strain mice. It was found that while a significant higher proportion of the female mice could be classified as high CRIA producers in responses to BA-Ar, no difference could be demonstrated if the inducing antigen was KLH-Ar. It is proposed that the difference observed with the two antigens may be due to the more selective activation by KLH-Ar of a small subset of high affinity Ar-specific clones--which may be enriched for CRIA + precursors--in both normal and immune defective mice. In contrast, BA-Ar may 'sample' more of the total anti-Ar repertoire and thus reveal within it an xid-determined depression in the proportion of CRIA + clones. Finally, it is noted that the influence of xid appears to be largely of a stochastic, and not an absolute character.

A strong delayed-type footpad reaction was established in mice using azobenzenearsonate-acetyl bovine serum albumin (ABA-AcBSA) as an antigen. Male ddY/S mice were sensitized by subcutaneous injection of 100 microliter of Freund's complete adjuvant - saline (1:1) emulsion containing 100 microgram of the antigen and challenged by subcutaneous injection of 2.5 microliter of Freund's incomplete adjuvant - saline (1:1) emulsion with 2.5 microgram antigen in the footpad on the 10th day after the sensitization. The analysis of the system which fulfilled criteria for delayed-type hypersensitivity with regard to kinetics, passive transfer and histology of the footpad reaction, and the effect of dexamethasone, indomethacin and quinacrine on the footpad reaction were described.

It has been firmly established that helper T lymphocytes respond to antigen only when it is presented in the context of class II major histocompatibility complex molecules. This has fostered the concept that the nominal antigen becomes physically associated with class II molecules on antigen-presenting cells (APCs). However, the structural components of antigen molecules that actually interact with class II molecules on APCs (agretopes) and with antigen receptors of T lymphocytes (epitopes) have not been precisely defined. This question was addressed using murine T cell clones specific for the model synthetic antigen L-tyrosine-p-azobenzenearsonate (ABA-tyr), which induces T cell responses despite its small size and simple structure, and a series of analogues of the homologous immunogen. Two experimental approaches were taken. First, APCs were pulsed with analogues and used to stimulate T cell proliferation. The patterns of stimulation segregated the clones into two specificity groups and indicated that the epitope recognized by the T cell included the arsonate group and elements of the side chain of tyrosine. Second, analogues that failed to stimulate were used to block the presentation of ABA-tyr in an effort to define the agretope. The blocking protocol involved serially pulsing APCs with analogue and with ABA-tyr prior to culturing with T cells. Compounds containing the core azo-linked ring structure blocked presentation of ABA-tyr in a dose-dependent fashion, whereas p-arsanilic acid and L-tyrosine were ineffective. The blocking was specific inasmuch as the compounds had no effect on the antigen-induced proliferative responses of unrelated T cell clones.(ABSTRACT TRUNCATED AT 250 WORDS)

Chemotactic activity at the site of an allergic air-pouch inflammation induced with azobenzene-arsonate-conjugated acetyl bovine serum albumin as an antigen was studied and a close correlation of the chemotactic activity with the intensity of leukocyte migration was demonstrated. In the period of vigorous leukocyte immigration into the fluid in the allergic air-pouch, chemotactic activity of the exudate was maintained at a high level, while no significant activity was detected after the number of leukocytes in the pouch fluid reached maximum.

Long-term cultures of functional T cells were grown from mice injected with azobenzene arsonate (ABA) coupled spleen cells or mice painted with the diazonium salt of p-phenylarsonate. All cell lines produced delayed hypersensitivity to ABA. When tested for helper function in an antibody-forming cell assay only one line, AA3, helped plaque-forming cell formation. It was able to help anti-FLU response of FLU-HGG primed B cells to ABA-KLH-FLU. The help required linkage of the ABA and FLU determinants because cultures stimulated with a mixture of ABA-KLH and TNP-KLH-FLU did not show increased PFC. These experiments demonstrate hapten-specific help in a system which eliminates involvement of passively absorbed antibody in hapten recognition.

T-cell antigen binding molecules (TABM) specific for trinitrophenol (TNP), oxazalone, azobenzenearsonate or sheep erythrocytes were purified by affinity to antigen, adsorption to monoclonal antibodies to antigen binding molecules or were synthesized by translation of immunopurified mRNA for TABM in vitro. These molecules and a T-cell line, BW5147, membrane protein bound by rabbit antibodies to TABM were radiolabeled by 125I, digested with Staphylococcus V8 protease, and peptides of the proteolytic digest were resolved by 2D-gel peptide mapping. Comparison of the peptide maps of these proteins and amino acid analysis of T-cell antigen binding molecules specific for TNP or sheep erythrocytes indicate similarities and distinctions suggesting variable and constant domains in these molecules.