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J W Goodman

Publications and source records attributed to J W Goodman.

At least 19 recordsLinked to original sources
PubMed1989

Arsonate-specific murine T cell clones. V. Antigen presentation by L cells transfected with normal and mutant class II genes.

Class II-restricted murine T cell clones specific for the immunogenic determinant L-tyrosine-p-azobenzenearsonate failed to proliferate to Ag presented by L cell lines transfected with and expressing the appropriate class II genes, but are activated to kill the APC in an Ag-dependent, MHC-restricted manner. Inhibition of APC proliferation was used as an assay to determine the relative contributions of polymorphic sites on the class II alpha- and beta-chains to MHC-restricted activation of I-A beta k-restricted cloned T cells. Transfectants expressing A beta k in conjunction with the alpha chain of k, u, or d were equally effective APCs, whereas transfectants expressing A beta u were completely ineffective, implicating the beta-chain as more critical for the presentation of L-tyrosine-p-azobenzenearsonate. Site-directed mutagenesis of polymorphic positions in the beta chain revealed a remarkable stringency for the k haplotype, in contrast to the relaxed alpha-chain requirement. These results, in conjunction with others, indicate that the relative contribution of polymorphic sites on class II alpha- and beta-chains to T cell Ag recognition can differ markedly, and, furthermore, may vary as a function of the Ag.

Animals↗
PubMed1988

Production of a fibronectin-associated lymphokine by cloned mouse T cells.

Azobenzenearsonate-specific cloned mouse T cells able to transfer delayed hypersensitivity reactions in vivo produced macrophage agglutination factor (MaggF) after stimulation with mitogen or antigen in vitro. Mitogen (Con A) elicited MAggF production directly from T cells. Responses to Ag were Ag-specific, required syngeneic accessory cells in addition to T cells, and were independent of T cell fine specificity for azobenzenearsonate. Mouse MAggF shared a number of biochemical and immunochemical properties with the fibronectins (FN): 1) high Mr similar to that of plasma FN; 2) binding to gelatin, heparin, and polyclonal antibodies and mAb specific for cellular and plasma FN; 3) inhibition of activity in solution by monoclonal anti-human FN directed against plasma FN gelatin-binding domain; and 4) action on peritoneal exudate macrophages mediated through a FN-receptor cross reactive with one on human monocytes. MAggF production required active protein synthesis and was associated with significant increases in gelatin-binding immunoreactive FN (Mr 440 kDa on immunoblotting) in culture supernatants and T cell lysates. Metabolically labeled peptides could be precipitated by anti-FN from culture supernatants of activated T cells. Stimulated cultures contained significantly more cells with immunohistologically demonstrable cytoplasmic FN than unstimulated control cultures. We suggest that T cell FN is a distinct species of cellular FN which may play an important role in mediating delayed hypersensitivity inflammatory reactions in vivo.

Animals↗
PubMed1988

The murine T-lymphocyte response to tyrosine-azobenzenearsonate. Characteristics of a low responder haplotype T-cell clone.

An I-Ab-restricted, L3T4+ Ly2- T-cell clone, 5R-4F3, specific for ABAtyr was established in culture from a B10.A(5R) mouse. Since b haplotype mice respond weakly to ABAtyr compared to other haplotypes, this is a candidate clone of low responder phenotype. In support of this contention, 5R-4F3 grew very poorly under conditions that supported the vigorous growth of E beta bE alpha k-restricted T-cell clones from the same mouse. The I-A (low responder) and I-E (high responder) restricted T-cell clones also differed in their responses to apc pre-pulsed with antigen, compared to apc with antigen present continuously during culture. The low and high responder clones responded comparably to IL-2. Attempts to elevate the response of C57BL/6 mice to ABAtyr in vivo by injecting them with human recombinant IL-2 and antigen together were only partially successful: C57BL/6 mice treated in this way showed a 3-5-fold increase in their proliferative responses to ABAtyr, which was at best only one quarter of the level of response shown by high responder A/J mice to the same antigen dose.

Animals↗
PubMed1987

Arsonate-specific murine T cell clones. IV. Properties of I-E- and I-A-restricted clones.

The T cell antigen L-tyrosine-p-azobenzenearsonate is unique in being a simple determinant that can be presented in the context of both I-A and I-E. I-E-restricted T cell clones derived from B10.A(5R) mice were found to fall into three groups: Type I clones recognized antigen only in the context of syngeneic apcs, Type II clones recognized antigen with the same highly specific major histocompatibility complex restriction but in addition proliferated in response to allogeneic stimuli; Type III clones were "degenerate" in their major histocompatibility complex-restricted recognition of antigen and proliferated when antigen-presenting cells bearing Eb beta Ek alpha (syngeneic), Ek beta Ek alpha, or Ed beta Ed alpha were used. These observations allow some conclusions to be drawn about sites on the I-E molecule that may be functionally significant in the presentation of this antigen. By using the B cell hybridoma LK35.2 as target cells, some of these T cell clones act as cytotoxic cells in the Class II-restricted manner predicted from the results of proliferative assays. Class II-restricted cytotoxicity can therefore be controlled by both I-A and I-E mouse Ir gene loci.

Animals↗
PubMed1986

Arsonate-specific murine T cell clones. III. Correlation between clonotype expression and fine specificity for analogs of L-tyrosine-p-azobenzenearsonate.

Despite recent advances in our understanding of T cell antigen receptor structure, relatively little is known about the role of this receptor in MHC-restricted antigen recognition. To study this problem, we have developed a panel of ABA-Tyr-reactive, I-Ak-restricted T cell clones that differ in their ability to recognize structural analogs of ABA-Tyr. Three fine specificity groups have been defined. In each group, ABA-Tyr elicited the strongest response of any of the antigens tested. Group I clones responded to ABA-conjugated hydroxyphenyl-ethanol (ABA-HPE). Group II clones responded to ABA-conjugated hydroxyphenyl-methanol (ABA-HPM) but not to ABA-HPE, and group III clones responded only to ABA-Tyr. These studies show that differences as small as a single methylene group can dramatically affect fine specificity. Because these clones are all I-Ak-restricted, it was possible to correlate receptor serology with fine specificity. To this end, monoclonal anti-clonotypes were made against clone 16-F2 from group I and used to study the relationship between fine specificity and clonotype expression. A panel of 15 T cell clones studied with four anti-clonotype antibodies showed a strict correlation between clonotype expression and fine specificity. Taken together, these data suggest that the structure recognized by the anti-clonotype antibodies is a determinant of receptor fine specificity.

Animals↗
PubMed1986

Functional substructure of antigen molecules.

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)

Animals↗
PubMed1985

Idiotypic properties of the murine anti-arsonate antibody response: B- and T-cell influences.

In a previous report characterizing the arsonate (ABA)-specific plaque-forming cell (PFC) responses of A/J mice induced by ABA-KLH, two interesting characteristics of the idiotypic (Id) profile were noted: (1) an apparent Id selectivity in the isotype switch since the earliest appearing IgG PFC in the primary response were significantly more "cross-reactive Id" (CRI)-dominant than the IgM PFC population, and, (2) a temporal waning of CRI dominance with time among IgG PFC, from 75-100% CRI+ PFC to about 25-45% CRI+ PFC in secondary responses. Experiments were performed to determine whether these effects are largely attributable to T or to B cells. Mice were immunized with a T-independent (TI) form of ABA (ABA-Brucella abortus) and apparent Id selectivity was observed; the earliest IgG PFC averaged 75% CRI+ while IgM PFC were only 39% CRI+. Due to the TI nature of the Ag, this provides suggestive, but not conclusive, evidence that the Id asymmetry in the isotype switch may be attributable to the direct interaction of Ag with B cells. Other studies addressed the temporal shift in CRI dominance. First, it was found that preexposure of mice to either KLH or to ABA (on an irrelevant carrier) resulted in diminished CRI dominance in subsequent "primary" responses to ABA-KLH. Secondly, adoptive transfer experiments with B and T cells from virgin mice (Bv, Tv) or ABA-KLH-primed mice (Bp, Tp) showed that recipients of Bv + Tp or Bp + Tv generated anti-ABA PFC responses with intermediate CRI levels. The Tv cells had some preferential tendency to activate CRI+ clones in the Bp population. The results demonstrate that CRI levels are jointly determined by the immune status of both B and T cells. A simple model is offered which accounts for early Id dominance and its gradual decline and has as its central postulate the assumption that CRI+ B cells in the virgin ABA-specific repertoire have an affinity advantage over CRI- clones.

Animals↗
PubMed1985

Interleukin 3 promotes erythroid burst formation in "serum-free" cultures without detectable erythropoietin.

Erythroid burst-forming units (BFU-E) from mouse bone marrow were grown for 7 days in agar or serum-free methylcellulose cultures in the presence or absence of erythropoietin (Ep) and/or interleukin 3 (IL-3). It was found that IL-3, even in the absence of serum and detectable Ep, was able to stimulate the full development of many erythroid bursts. This IL-3 effect was cell-dose dependent and did not appear to correlate with Ep dose. Spontaneous bursts and those stimulated by Ep only were rare and when seen were very small relative to those produced by IL-3 or IL-3 plus Ep. When addition of IL-3 or Ep to 7-day cultures was delayed, IL-3 but not Ep was shown to maintain BFU-E. No evidence was found by radioimmunoassay that Ep was produced or released in 7-day, "serum-free" cultures of bone marrow nor was Ep activity detected in culture media except those to which it had been added deliberately.

Animals↗
PubMed1985

Arsonate-specific murine T cell clones. II. Delayed-type hypersensitivity induced by P-azobenzenearsonate-L-tyrosine (ABA-Tyr).

To study T cell idiotype expression at the functional level, we developed a hapten-specific delayed-type hypersensitivity (DTH) system by which we avoid the complication of anti-hapten antibody and which is specific only for the immunizing hapten, and not for conjugate specific determinants. Immunization with ABA-Tyr and challenge with ABA diazonium induced footpad swelling with the characteristics of DTH. Anti-ABA antibodies did not contribute to this reaction, as they were undetectable in mice immunized with ABA-Tyr. Furthermore, this ABA-Tyr-specific DTH was under Ir gene control identical to that reported for ABA-Tyr-specific lymphocyte proliferation. All mouse strains tested responded to ABA-Tyr except those of the b haplotype across the entire Ia region. In contrast, contact sensitivity induced by ABA diazonium was not under apparent Ir gene control, probably reflecting 1) different specificities of the induced T cells and 2) the production of anti-ABA antibodies that contribute to the footpad swelling via an Arthus reaction. Having shown that ABA-Tyr can induce T cells mediating DTH, we then examined ABA-Tyr-reactive T cell clones, propagated in vitro, for their ability to mediate DTH. Such clones elicited a response identical to that seen with in vivo immunization with respect to dose dependency, I-Ak restriction, and antigen specificity.

Animals↗
PubMed1984

Differential induction of help and suppression in mice by bifunctional antigens administered via different routes.

Bifunctional antigens composed of one L-tyrosine-p-azobenzenearsonate (Tyr-ABA) carrier epitope and one dinitrophenyl (DNP) haptenic epitope separated by 6-aminocaproyl or polyprolyl spacers induced weak IgM anti-DNP plaque-forming cell (PFC) responses in the spleens of mice immunized intraperitoneally, without detectable IgG PFC. However, the same antigens introduced into the footpads induced IgG PFC responses in the draining lymph nodes which rose to levels greater than 100/10(6) viable lymphocytes. Moreover, the response in the lymph nodes to booster injections of antigen was characteristic of secondary T-dependent antibody responses, whereas the splenic secondary response simply mirrored the primary. The magnitude of the IgG PFC response was influenced by the size of the spacer and by the strain of mice, although genetic control did not map to the major histocompatibility complex. Prior i.p. immunization suppressed the IgG response to subsequent immunization in the footpads. This suppression could be transferred to normal syngeneic recipient mice with spleen cells from suppressed donors. Suppressor activity was eliminated by treating the spleen cells with anti-Thy-1 antibody prior to transfer, establishing the T-cell dependency of suppression. Suppression was also induced by Tyr-ABA itself, but not by DNP-lysine, indicating the epitope specificity of the suppressor cells. Thus, bifunctional antigens induce dominant suppression in the spleen but significant help in lymph nodes.

Animals↗
PubMed1984

Immunogenicity of biotinylated hapten-avidin complexes.

The efficacy of avidin as a carrier for the generation of anti-hapten antibodies was assessed in mice by immunization with complexes of avidin and synthetic peptides containing biotin and an epsilon-dinitrophenyl (DNP) lysine residue. The synthetic haptens were constructed with 0, 1 or 2 6-aminocaproyl groups as spacers between the biotin and DNP-lysine moieties. Complexes without a spacer did not induce anti-DNP antibody responses, while those with two spacers induced stronger responses than those with only one spacer. However, the anti-DNP responses to avidin-biotinylated hapten complexes were considerably weaker than responses to a conventional hapten-protein conjugate (DNP-ovalbumin), and, like "T-independent" antigens, failed to induce significant immunological memory. The distribution of isotypes in the anti-DNP antibodies produced to avidin-biotin-6-aminocaproyl-epsilon-DNP-lysine-alanine and DNP-ovalbumin was similar, but the former antigen induced significantly lower levels of antibody in (CBA/N X BALB/c) F1 male mice with the xid defect than in phenotypically normal female littermates, and also induced significant responses in nu/nu mice, in contrast to DNP-ovalbumin. These findings suggest that there is at least a "T-independent" or "T-efficient" component in the response to avidin-biotin complexes, perhaps due to the tetrameric structure of the molecule. Estimates of the depth of the receptor site for biotin were obtained by using the complexes to competitively inhibit the binding of anti-DNP antibody to plates coated with DNP-protein. The findings were consonant with the data on immunogenicity (capacity to induce anti-DNP antibody responses) and suggested that the receptor site has a depth of 16-26 A.

Alanine↗
PubMed1984

The anatomy of an antigen molecule: functional subregions of L-tyrosine-p-azobenzenearsonate.

The structural components of antigen molecules that interact with class II major histocompability complex (MHC) molecules on antigen-presenting cells (APCs) (agretopes) and with antigen receptors of T-lymphocytes (epitopes) in class II restricted T-cell responses have not been precisely defined. This issue was addressed here using murine T-cell clones specific for the simple immunogen L-tyrosine-p-azobenzenearsonate (ABA-tyr) and a series of analogs of the homologous antigen. Two experimental approaches were used. First, APCs were pulsed with analogs 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 in the side chain of tyrosine. Furthermore, the clones manifest different sensitivities to antigen. Second, non-stimulatory analogs were used to block the presentation of ABA-tyr in an effort to define the agretope. Compounds containing the azophenyl group 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 giant keyhole limpet hemocyanin (KLH) or hen egg white lysozyme (HEL)-reactive T-cell clones. The blocking pattern indicated that the feature required for productive association with the APC centered on the planar structure of the azo-linked aromatic rings, with little or no contribution from either the arsonate moiety or the tyrosyl side chain. We propose that this structure forms an agretope for this family of compounds.

Animals↗
PubMed1984

In vitro studies of erythropoietic progenitors (CFU-E) in marrow from neonatal and young mice.

Reports that cells from neonatal animals produce CFU-E in response to erythropoietin (Ep) at doses lower than are required by adults indicated that young mice would provide a good system to study the hormone response of erythropoietic progenitors. Studies presented here confirm the increased Ep responsiveness until 3-4 weeks of age reported previously by others. Along with the enhancement in their Ep responsiveness, bone marrow cells from young animals produce many more CFU-E per 10(5) cells plated than do adult animals; i.e., neonates have a higher plating efficiency for CFU-E in the presence of erythropoietin. Neonatal animals, in contrast to adults, contain in addition erythropoietic progenitors in their bone marrow capable of growth in vitro without added hormone. Because of the large numbers of "endogenous" colonies, a study was done to explore the possibility that a small amount of Ep was present in the culture medium. Antiserum to erythropoietin was added directly to cultures in another study to remove any undetected hormone. The results indicate that CFU-E from neonatal mouse bone marrow can develop in culture in the absence of detectable Ep.

Aging↗
PubMed1983

Idiotype profile of an immune response. II. Reversal of the relative dominance of major and minor cross-reactive idiotypes in arsonate-specific T-independent responses.

Two different cross-reactive idiotype (CRI) groups are distinguishable in the Ab response of A/J mice to the p-azobenzenearsonate (ABA) hapten: CRIA and CRIm. These two groups showed distinct patterns of relative dominance in the ensuing response depending on whether the inducing Ag was a T cell-dependent (TD) form of ABA, such as ABA-KLH or ABA-CGG, or a T-independent type 1 (TI-1) form, such as ABA-Brucella abortus or ABA-lipopolysaccharide (LPS), and on whether the response was elicited in vivo or in vitro. The CRI+ component of primary in vivo plaque-forming cell (PFC) responses to TD ABA Ags was largely (greater than 90%) CRIA+ as was, to a slightly lesser extent (greater than 75%) the CRI+ portion of secondary or hyperimmune serum Ab or PFC responses to the same Ags. In contrast, in vivo primary and hyperimmune PFC responses to ABA-Bru or ABA-LPS showed a significantly lower CRIA/CRI ratio, averaging 0.5-0.6, with some individual mice giving figures as low as 0.2, indicating predominance of CRIm over CRIA. Serological analysis of hyperimmune anti-ABA Abs from a group of 5 A/J mice immunized with ABA-Bru gave a figure of less than 0.5 for the CRIA/CRI ratio. The most striking disparity from the TD pattern was seen in primary in vitro PFC responses to the TI ABA Ags; here ratios of less than 0.2 were generally seen. Since T cell removal did not alter the Id pattern in the TI responses, CRIA-specific Ts cells do not account for the weak expression of CRIA in such responses. We propose a model that explains these results on the basis of differential expression of IdX dominance by two distinct B cell subpopulations--equatable to the Lyb-5+ and Lyb-5- B cell subsets--along with differential relative activation of these subsets in different types of responses. Examination of anti-ABA PFC responses of F1 progeny of CBA/N and A/J mice to ABA-Bru lends support to this hypothesis since CRIA expression was significantly lower in mice with the xid defect.

Animals↗
PubMed1983

Arsonate-specific murine T cell clones. I. Genetic control and antigen specificity.

The antigen-induced proliferative response of lymph node cells (LNC) from mice sensitized to the monofunctional antigen L-tyrosine-p-azobenzenearsonate (ABA-Tyr) was used to monitor genetic control. All strains tested mounted significant responses, but those that were H-2(b) at both the I-A and I-E loci [B10., B6., B10.A(18R), A.BY, and C3H.SW] gave consistently weaker responses than other haplotypes. The F(1) progeny of matings between high and low responder phenotype parents (DBA/2 and B6, respectively) were high responders, establishing the dominance of the responder trait. Proliferative responses of LNC to ABA-Tyr were blocked by the appropriate anti-Ia monoclonal reagents. For example, B10.A(4R) LNCI (I-A(k), I-E(b)) were blocked by anti-I-A(k), whereas B10.A(3R) LNC (I-A(b), I-E(k)) were blocked by anti-I-E(k). Long-term cultures of T cell lines specifically reactive to ABA-Tyr were established from LNC of A/J mice immunized with ABA-Tyr and were cloned by limiting dilution. The proliferative responses to ABA-Tyr of 14 out of 15 clones tested were I-A restricted on the basis of activation by antigen-presenting cells from appropriate recombinant strains and the blocking activity of the monoclonal anti-Ia antibodies. The response of the other clone was I-E restricted. The fine antigen specificity of the clones was studied using structural analogs of the homologous antigen to induce proliferation. The clones could be divided into three types with respect to responsiveness to ABA-histidine (ABA-His). One group responded about equally well to ABA-His and ABA-Tyr. A second set responded less strongly to ABA-His than to ABA-Tyr, while the third showed no response above background to ABA- His. In all instances, the ABA-His-responding clones discriminated exquisitely between the 2-azo and 4-azo histidine isomers, responding only to the 4-azo compound. These T cell clones provide extremely useful tools for studies of T cell specificity, antigen recognition and lymphoid cell interaction systems.

Animals↗