cAMP-mediated suppression of a Th1 clone associated with an alteration of the intracellular redox environment.

We have shown previously that the elevation of intracellular cAMP in antigen or anti-CD3-activated murine Th1 clones in the absence of antigen inhibits antigen-induced proliferation and the production of IL-2 by H2O2-mediated oxidation of p56lck and inhibits antigen-induced production of interferon-gamma by the induction of intracellular nitric oxide. Moreover, activated Th1 clones are resistant to cAMP-induced suppression. These results suggest that the immunosuppression of Th1 cells mediated by elevated intracellular cAMP is associated with an alteration in the intracellular oxidation/reduction environment. Here we report that the culture of an antigen or anti-CD3-activated murine Th1 clone with the adenylcyclase agonist forskolin (FSK) in the absence of antigen reduces the activity of intracellular catalase, and diminishes levels of intracellular reduced glutathione (GSH). Resting cells resistant to cAMP-induced suppression have higher intracellular GSH levels than antigen-activated cells susceptible to cAMP-induced suppression. The results provide further evidence that cAMP-induced suppression of Th1 clones is mediated by profound alterations in the intracellular redox environment and may be used to selectively inactivate Th1 cells activated by antigen.

Differential regulation of T cell receptor-mediated Th1 cell IFN-gamma production and proliferation by divergent cAMP-mediated redox pathways.

Culture of an H-2(s)-restricted, bovine myelin basic protein (BMBP)-specific murine Th1 clone with the adenyl cyclase agonist forskolin (FSK) or isobutylmethylxanthine (IBMX), an inhibitor of cAMP catabolism, before culture with anti-CD3 or BMBP and antigen-presenting cells (APC) suppressed antigen or anti-CD3-induced proliferation and production of interferon-gamma (IFN-gamma). Other H-2(s)-derived or H-2(b)-derived clones specific for BMBP or keyhole limpet hemocyanin (KLH) were similarly affected. FSK did not affect the expression of CD4 or the T cell receptor (TCR) but did diminish levels of the phosphorylated (activated) mitogen-activated protein (MAP) kinases early response kinase-1 (ERK-1) and ERK-2. Immunoblotting of lysates from an FSK-treated Th1 clone with antibodies to a carboxy-terminal epitope of p56(lck), a signal transduction enzyme upstream from ERK-1 and ERK2, did not detect p56(lck) unless the lysates were reduced prior to electrophoresis. Immunoblotting of nonreduced lysates with antibodies to an amino-terminal epitope demonstrated p56(lck) with a lower apparent molecular weight, characteristic of oxidized proteins. Reduction restored the detection of p56(lck) by anticarboxy-terminal p56(lck) and to mobilities indistinguishable from controls detected by the antiamino-terminal p56(lck). N-acetylcysteine or catalase prevented FSK-induced suppression of antigen-induced proliferation and the loss of carboxy-terminal epitopes of p56(lck). An inhibitor of cAMP-dependent protein kinase A (PKA) or nitric oxide synthase (NOS) did not affect FSK-induced inhibition of antigen-induced proliferation. In contrast, inhibitors of PKA or NOS, but not catalase, prevented FSK-induced suppression of IFN-gamma production. Moreover, immunoblots of lysates precipitated with anti-p56(lck), phosphotyrosine, or CD4 demonstrated that in FSK-treated, anti-CD3-stimulated cells, p56(lck) is not associated with CD4 zeta chain, nor is p56(lck) or zeta chain phosphorylated. In vitro kinase assays demonstrated that p56(lck) from FSK-treated cells does not have kinase activity. Taken together, the results suggest that an elevation of intracellular cAMP (in the absence of antigen) creates an oxidative environment that oxidizes and inactivates p56(lck) by an H(2)O(2)-dependent, PKA-independent mechanism and inhibits the production of IFN-gamma by an NO, PKA-dependent mechanism. Thus, antigen-induced proliferation and IFN-gamma production in a Th1 clone are controlled separately by different cAMP-dependent, redox-based mechanisms.

Production of serum immunoglobulins and T cell antigen binding molecules specific for cow's milk antigens in adults intolerant to cow's milk.

The immune response to three cow's milk antigens, beta-lactoglobulin (BLG), alpha-lactalbumin (AL), and casein (CA) was studied in 15 milk-intolerant adult patients and 11 adult controls. IgG, IgE, and IgG subclasses (IgG1, IgG2, IgG3, IgG4) and T cell-derived antigen-binding molecules (TABM) specific for each antigen were measured in both groups. In the patient group, a significant elevation of total IgG and TABM against each of the milk antigens was found as well as raised levels of IgG1 to BLG and CA, IgG4 to BLG, and IgE to CA. TABM specific for BLG were isolated by affinity for BLG and found to be Mr 28,000-46,000 polypeptides functionally and physically associated with TGF-beta1 and TGF-beta2. These results indicate a Th2-type immune response to the milk antigens in milk-intolerant individuals compared with the control group which shows a pattern typical of anergy or deletion.

Elevation of intracellular cyclic AMP induces an anergic-like state in Th1 clones.

Because elevated intracellular cAMP suppresses T cell receptor (TCR)-mediated effector activity and/or proliferation in response to antigen but does not always affect IL-2-stimulated proliferation, the effects of cAMP on a T lymphocyte response to antigen resemble antigen-induced anergy. To test the hypothesis that elevated cAMP induces anergy in T lymphocytes, we have precultured murine Th1 clones responsive to porcine myelin basic protein (PMBP) with dibutyryl cyclic AMP (dbcAMP) or forskolin and subsequently removed the dbcAMP or forskolin and measured the proliferative response of the clones to antigen and antigen-presenting cells (APC) in the presence or absence of exogenously added interleukin-2 (IL-2). Cells precultured with dbcAMP or forskolin for 3 days did not proliferate or produce IL-2 in response to antigen and APC, but did proliferate to antigen and APC in the presence of IL-2. Cells that had not been stimulated recently with antigen/APC or IL-2 were not affected by dbcAMP, while cells stimulated recently with antigen/APC and IL-2 were susceptible to the anergizing effect of dbcAMP. These observations support the hypothesis that elevation in intracellular cAMP in antigen-activated Th1 clones, prior to subsequent culture with antigen, induces a state of anergy.

Intracameral injection of antigen potentiates the production of antigen-specific T cell proteins in serum after the induction of delayed-type hypersensitivity.

PURPOSE: To determine whether the introduction of antigen into the anterior chamber induces the production of extracellular antigen-specific T cell proteins (T cell antigen-binding molecules [TABM] specific for the antigen. METHODS: Balb/c mice received an intracameral or subconjunctival injection of trinitrophenylated spleen cells (TNP spleen cells) before skin sensitization and challenge with picrychloride. The production of TNP-specific TABM in serum was quantified by enzyme-linked immunosorbent assay-based antigen binding and immunoblotting using a rabbit antiserum raised against a monoclonal antigen-specific T cell protein that induces suppressor T cells. RESULTS: Intracameral, but not subconjunctival, injection of TNP spleen cells before contact sensitization increased the level of TNP-specific TABM in serum. At 2 days, TABM levels began to rise and peaked at 5 days after contact sensitization. Anterior chamber injection of TNP spleen cells alone into nonimmunized mice did not induce a detectable increase of TNP-specific TABM in serum. Mice that did not receive an intracameral TNP spleen cell injection but were made contact sensitive to TNP showed a diminished TNP-specific TABM response. Specificity of the TABM induced by intracameral-injection and sensitization for the TNP hapten was documented by showing that the induced TABM binds TNP-bovine serum antigen (BSA) but does not bind either BSA alone or azobenzenearsonate-ovalbumin and that immune sera raised against ovalbumin did not contain an increase in TNP-specific TABM. TNP-specific TABM in the sera of mice receiving an intracameral injection of TNP spleen cells, followed by contact sensitization with picrylchloride (PCl), were purified by affinity for TNP and were resolved by polyacrylamide gel electrophoresis and immunoblotting as M(r) 110,000 polypeptides. CONCLUSIONS: Serum levels of TNP-specific TABM induced by contact sensitization alone are enhanced fourfold to fivefold when sensitization is preceded by the intracameral, but not the subconjunctival, injection of TNP spleen cells. The authors propose that besides suppression of systemic delayed type hypersensitivity, the intracameral injection of antigen into nonimmune mice may prime an increased production of TABM, which may indicate systemic activation of the immunoregulatory T cell circuit.

Increased basal levels of free plasminogen activator activity found in human aqueous humor.

PURPOSE: To quantify the basal free t-PA activity in human aqueous humor. METHODS: Human aqueous humor obtained by simplified pipette paracentesis at cataract surgery was tested for free t-PA activity in a revised 15-hour amidolytic assay using a t-PA standard curve (n = 15). Total antigenic levels of PAI-1, the principal PA inhibitor, were determined using an ELISA kit. The available PAI-1 activity was tested indirectly using anti-human PAI-1 antibody blocking before t-PA activity assay (n = 11). Plasminogen activator type was determined by anti-human t-PA and urokinase (u-PA) antibody blocking before activity assay (n = 7). RESULTS: Free PA activity ranged widely (0.072 to 0.47 IU/ml; mean, 0.20 +/- 0.10 IU/ml) and was almost completed inactivated (> or = 89%) by antibody against human t-PA but not by the u-PA antibody. PAI-1 total antigen also ranged widely between (0.25 to 8.0 ng/ml; mean, 2.25 +/- 2.54 ng/ml). However, pretreatment of samples with PAI-1 antibody or by acidification (pH 3.2) to inactivate inhibitors did not increase t-PA activity levels. CONCLUSIONS: A basal-free t-PA activity, which is predominantly t-PA, is present in human aqueous humor at approximately 20 times higher levels than previously found with an earlier assay. This activity is estimated to represent roughly 10% of total released t-PA antigen. PAI-1 is present in aqueous humor at levels considerably lower than reported values for plasma in a predominantly PA-complexed or inactive form.

T cell-derived antigen-specific humoral immune response. II. Further characterization of the response and the antigen binding T cell immunoproteins.

Murine T lymphocyte-derived proteins which bind nominal antigen specifically (TABM) were detected in serum during a humoral immune response to bovine serum albumin. The TABM response was observed only when the adjuvants polyadenylic:polyuridylic acid complex (poly(A: U)), Freund's complete adjuvant or Titermax were used concurrently with antigen to immunize. When poly(A:U) was used, higher doses of antigen were optimal to stimulate TABM production than those required to stimulate immunoglobulin production. The binding of TABM in the serum of BSA-immunized mice to BSA in solid phase was inhibited specifically by 10-fold more BSA than that required to inhibit BSA-specific immunoglobulins suggesting that TABM have much less affinity for antigen than immunoglobulins. Immunoblotting of TABM in serum from BSA-immune mice, which bind BSA, demonstrated that these serum TABM are comprised of M(r) 110,000 polypeptides linked by disulfide bonds. Antigen-specific proteins in a lysate of an NP-specific T cell hybrid inhibited the recognition of serum TABM by anti-TABM antiserum while a lysate of a B cell hybridoma was not inhibitory, and serum TABM were adsorbed by monoclonal antibodies to TCR-C beta. These results provide more evidence that serum TABM may be a soluble analogue of the T cell receptor for antigen.

Tissue plasminogen activator activity in human aqueous humor.

PURPOSE: To determine the levels of free plasminogen activator activity in human aqueous humor and to identify the type of activity (i.e., tissue-type t-PA or urokinase-type u-PA) that is responsible. METHODS: Aqueous humor was obtained by a simplified pipette paracentesis before cataract surgery in 31 subjects, ages 57 to 93 years. Levels of plasminogen activator activity were determined using a modified 17-hour specific amidolytic assay. The type of plasminogen activator was investigated in selected samples based on its dependence on soluble fibrin, inhibition by amiloride, and specific antibody blocking. Activity-antigen ratios were compared in seven samples. RESULTS: Plasminogen activator (PA) activity was present in all samples tested. PA activity ranged widely between 0.54 and 26.7 mIU/ml, with a mean value of 10.8 +/- 8.1 mIU/ml. Soluble fibrin, a known stimulator of tissue-type plasminogen activator (t-PA), was required in the assay system. Its absence decreased the measured activity by more than 90%. Amiloride, a known inhibitor of urokinase-type PA, had little or no effect in selected samples tested. The activity was blocked by anti-human t-PA antibodies but not by antibodies against human u-PA, further defining the type of PA responsible for the detected activity. t-PA antigen levels showed less variation among individuals than did activity levels. Antigen-activity ratios ranged between 89 and 552. CONCLUSION: Plasminogen activator activity is present in the human aqueous humor in measurable quantities. The type of PA activity present is almost exclusively t-PA. t-PA activity varies more widely than antigen, as is the case in plasma.

Partial amino acid sequence of monoclonal extracellular antigen-specific T cell proteins.

Antigen-specific molecules secreted by murine T cell hybrids (TABM) specific for 4-hydroxy 3-nitrophenyl (NP) or azobenzenearsonate (ABA) were purified from ascitic fluid by ion exchange chromatography and/or affinity for antigen. Partial amino acid sequence of reduced Mr 72,000 NP-specific polypeptides and Mr 20,000 peptides prepared by treatment of the ABA-specific immunoprotein with cyanogen bromide was obtained and a septapeptide of the NP-specific TABM shared 3/7 residues with the ABA-specific TABM. Both TABM shared residues present in 95% T cell receptor for antigen (TCR) V alpha subgroup I and 83-96% murine immunoglobulin V kappa Fr3. These results provide evidence that extracellular antigen-specific T cell proteins are soluble analogues of TCR alpha chains and belong to the immunoglobulin supergene family.

Regulatory effect of interferon-gamma and phorbol esters on the surface expression and biosynthesis of MHC class I antigens by human leukemia cells.

We have used cell surface radioiodination, biosynthetic incorporation of [35S]methionine, and flow cytometry to analyze the effects of interferon gamma (IFN-gamma) and/or phorbol esters (PMA) on the turnover and expression of class I antigens of a human leukemia B cell line. Our results demonstrated that although both IFN-gamma and PMA enhance HLA expression, they act synergistically to increase by eightfold the amount of HLA polypeptides synthesized by the acute lymphoblastic leukemia cells and acted additively to augment the cell surface expression of HLA as quantified by flow cytometry. We observed a cyclic increase or decrease in the expression of class I antigens as a function of time in cell culture. IFN-gamma and/or PMA modulated this effect inducing more cells to express HLA maximally. These results suggest that there is a physiologic limit for the expression of major histocompatibility complex class I antigens.

Appearance of T lymphocyte-derived proteins specific for the immunizing antigen in serum during a humoral immune response.

Some T lymphocytes produce extracellular proteins that bind nominal antigen specifically (TABM), and these proteins exhibit potent immunoregulatory activity. We have utilized an ELISA for Ag binding by Ag-specific TABM to detect and quantitate the appearance of Ag-specific TABM in murine serum during a humoral Ir to protein Ag. The TABM response was specific for the inducing Ag, stronger and more rapid during a secondary response, and temporally distinct from the appearance of Ig. The non-Ig serum TABM were bound by mAb specific for TCR-alpha chains and isolated by affinity for Ag were Mr 110,000 polypeptides. The TABM response did not occur in scid/scid mice unless the mice were reconstituted with thymocytes and thymocyte-reconstituted scid/scid mice produced TABM, but did not produce Ig. The results suggest that soluble TABM are an Ag-specific humoral manifestation of the Ir of some T lymphocytes.

Specific antigen binding by proteins secreted by an antigen-specific T cell hybrid.

Antigen-specific molecules secreted by a murine T cell hybrid specific for azobenzene arsonate (ABA) were purified from ascites fluid by ion exchange chromatography and affinity for antigen. The antigen-specific proteins were purified 250 fold and were resolved predominantly as Mr 110,000 polypeptides by reduction and SDS-polyacrylamide gel electrophoresis. The ability of these molecules to bind antigen was analyzed by an ELISA using antigen-coated microtiter trays. Binding of the T cell proteins to antigen was detected with antisera specific for the proteins. Antigen binding to ABA-ovalbumin but not ovalbumin was optimal at 37 degrees C and protein derived from another T cell hybrid did not bind ABA-ovalbumin. Solid phase antigen binding was inhibited specifically by soluble ABA-ovalbumin, indicating that these T cell-derived proteins bind nominal antigen in the solid or liquid phase. It is suggested that these proteins represent a soluble, antigen specific manifestation of some T cell function.

T cell non-MHC-restricted antigen-binding molecules secreted or associated with the cell membrane are antigenically distinct.

Some T cells produce membrane-associated or soluble molecules which bind nominal antigen specifically (TABM) and effect immunoregulation or events similar to cell-mediated hypersensitivity. We have used polyclonal antisera raised against an azobenzene arsonate (ABA)-specific TABM secreted by an ABA-specific T cell hybrid or against TNP-specific polypeptides produced by immunoregulatory T cells to identify the expression of soluble (secreted) or membrane-associated TABM. Ascites fluid or culture medium containing a T cell hybrid or T cell lines, respectively, contain TABM recognized only by an antiserum specific for the secreted T cell hybrid (ABA-specific) derived TABM. Conversely, an antiserum that recognized the TNP-specific polypeptides detected cell-membrane associated TABM but did not bind TABM secreted by the T cell hybrid or cell lines.

T cell derived proteins from normal human sera and their relationship to T cell antigen binding molecules.

We have used procedures which have been developed to isolate murine T cell antigen binding molecules (TABM) in order to isolate TABM from normal human sera. To begin purification, ammonium sulfate (NH4)2SO4 was added to human serum and precipitated protein was dissolved in low salt buffer and resolved by ion-exchange chromatography on carboxymethylcellulose (CM). The most strongly CM nonadherent fraction was absorbed with anti-human albumin and anti-human immunoglobulin (Ig) antibodies conjugated to Sepharose beads. The resulting nonadsorbed 110,000, 70,000 and 45,000 Mr polypeptides were reactive in ELISA with a rabbit antiserum produced against non-Ig, anti-specific molecules of rhesus monkeys. These proteins possess alpha mobility upon immunoelectrophoresis and represent 0.02 to 0.05% of total serum protein. In addition, these proteins are bound by an antiserum made against a synthetic peptide corresponding to the J region of the TcR beta chain. We have made R28, a rabbit antiserum against these serum proteins which binds specifically to tetanus-specific polypeptides obtained from the culture supernatant of human T cell lines specific for tetanus. This antiserum also binds to proteins isolated from T cell but not B cell lines, and T cell proteins are able to inhibit the binding of R28 to the human serum polypeptides. The results suggest that the proteins isolated from normal human sera are T cell antigen binding molecules.

A monoclonal antigen-binding T cell immunoprotein: antigenic relatedness to T cell receptor beta chain FR1 V and J peptide segments: physicochemical distinctiveness from classical immunoglobulins and T cell receptor heterodimers.

The monoclonal murine T cell hybridoma, 51H7D, was previously shown to bind the arsazobenzene hapten and to produce a soluble antigen-binding molecule. In this paper we characterize this antigen-binding immunoprotein for its relationship to known T cell receptors serologically, using antibodies specific for variable region framework, or joining region peptides predicted from gene sequence and by biochemical means. The 51H7D cell expresses a protein with subunit size of approximately 31,000, that reacts antigenically with affinity-purified antibodies directed against synthetic first framework and joining segment peptides, corresponding to the gene sequence of the T cell receptor beta chain, YT35. This molecule does not react with affinity-purified antibodies directed against murine immunoglobulin, framework 1 sequences of alpha and gamma T cell receptors, or with antibodies against synthetic heavy chain joining segments. The subunit of mol. wt. 31,000 can form higher aggregates, notably in the mol. wt range of 60,000-70,000, depending upon extraction conditions. The soluble form of the antigen-binding molecule bears the J beta cross-reactive determinant and occurs predominantly as a charge restricted molecular species of approximate mol. wt 60,000-70,000. The purified molecule has a blocked N-terminus, but quantitative statistical analysis of its amino acid composition indicates a closer relatedness to T cell receptor beta chains and other antigen-binding T cell products, than it has to alpha, gamma or delta TCR chains. No evidence for more than one type of polypeptide chain was found and the polymerization is not dependent upon the formation of disulfide bonds. These studies raise the possibility that antigen-binding soluble T cell molecules might belong to a new family of immunoproteins, that is related to, but distinct from, classical immunoglobulins and alpha beta or gamma delta heterodimers.

Characterization of the molecules involved in thymocyte thymic epithelial cell adhesion.

Cell to cell adhesion is important for mechanisms of cellular recognition, growth, and differentiation. The identification of molecules involved in these interactions is necessary in order to understand the molecular basis of these processes. We have previously described the development of two different thymic epithelial cell lines (TECS and TECL). Using an assay with radiolabeled thymocytes we found that thymocytes can adhere specifically to these thymic epithelial cells. This adhesion is trypsin sensitive, suggesting that involvement of specific cell surface proteins. In the present study we further characterize and begin identification of the molecules involved in this interaction. We found that thymocyte binding to thymic epithelial cells requires the presence of Ca2+ and Mg2+ and is mediated by a molecule greater than 10,000 MW. Also, we identified several antibodies which inhibit the adhesion of thymocytes to TECS. The membrane nature of the molecule mediating this interaction was confirmed by the ability of thymocyte membranes to block the inhibitory antibodies.

Isolation of antigen-binding membrane molecules from an antigen-specific T-cell hybrid.

Cell surface-radioiodinated proteins of a murine T-cell hybrid specific for and able to bind azobenzenearsonate were isolated by adsorption to Sepharose beads conjugated with a rabbit antiserum to murine T-cell antigen-binding molecules. These isolated proteins, Mr 72,000, were found to bind specifically azobenzenearsonate while proteins isolated in this manner from the tumor parent BW5147 did not bind azobenzenearsonate. Similar cell surface proteins were isolated by affinity for antigen and immunoprecipitated with an antiserum to T-cell antigen-binding molecules. The results suggest that antigen-binding T cells express T-cell antigen-binding molecules as membrane receptors for antigen.

Phenotypic similarity between T-cell antigen binding molecules.

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.

Antigen-binding molecules of T-cells charge heterogeneity and structural lability.

T-cell products released by immune cells during culture and which bind specifically the nominal antigens, trinitrophenol (TNP) or oxazalone, were isolated from culture media by hapten-affinity chromatography and compared by isoelectric focusing and 2D-gel analysis. These proteins and an azobenzenearsonate-specific T-cell product synthesized in vitro by translation of mRNA from an azobenzenearsonate-specific T-cell hybrid were also compared for structural lability of the polypeptides. Polyclonal T-cell antigen-binding molecules (TABM) specific for TNP or oxazalone showed marked charge heterogeneity and distinctions in isoelectric focusing in an acidic pH gradient, while the azobenzenearsonate-specific, clonal T-cell product displayed restricted focusing. All TABM studied showed dissociation of Mr 70,000 polypeptides to Mr 45,000 and 25,000 polypeptides after treatment with guanidine. The results provide further evidence for distinctions and similarities between TABM.

Comparison of altered expression of histocompatibility antigens with altered immune function in murine spleen cells treated with ultraviolet radiation and/or TPA.

Previous studies in our laboratory demonstrated that several treatments that inhibited the ability of cells to stimulate the mixed lymphocyte reaction (MLR) also blocked the shedding of histocompatibility antigens and Ia antigens from murine spleen cells. In the present studies, one of these treatments, ultraviolet radiation (UV), was shown to cause an initial loss in the density of H-2K, IA, and IE antigens prior to the block in shedding observed after culture of these cells. Further analysis revealed that the UV-induced loss of antigens could be prevented by the presence of colchicine during irradiation. Biosynthetic analyses revealed the IA antigen synthesis was also inhibited in the UV-irradiated cells. Examination of the effects of a second agent, 12-0-tetradecanoylphorbol-13-acetate (TPA) on the turnover of histocompatibility antigens revealed that the biosynthesis and shedding of these antigens were accelerated by this agent. However, addition of TPA to UV-irradiated cells did not result in a reversal of the UV-induced block in biosynthesis of IA antigens. Results of immune function assays correlated with the biochemical studies: UV-irradiation inhibited the generation of the MLR, but TPA enhanced this reaction, and addition of TPA to mixed lymphocyte cultures with UV-irradiated stimulators did not reverse the UV-induced inhibition. These results suggest that, although the turnover of histocompatibility antigens may be affected by TPA and UV in an antagonistic fashion, additional factors other than the expression of histocompatibility antigens are operating in the inhibition of stimulation of an MLR by UV radiation or its enhancement by TPA.