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A particular phenol-formaldehyde resin of the resol type (P-F-R-2) was shown in a previous study to be a good tracer of patients with contact allergy to phenol-formaldehyde resins. Eleven patients with sensitivity to P-F-R-2 participated in the study in which chromatographically separated fractions of P-F-R-2 were patch tested. This procedure was repeated until a positive reaction to a single substance was noted. In this study 3 new sensitizers are reported. Investigations by mass spectrometry and nuclear magnetic resonance spectrometry identified the compounds to be 4,4'-dihydroxy-3-(hydroxymethyl)-diphenyl methane (4,4'-H-3-HPM), 4,4'-dihydroxy-3,3'-di-(hydroxymethyl)-diphenyl methane (4,4'-H-3,3'-HPM), and 4,4'-dihydroxy-3,5-di-(hydroxymethyl)-diphenyl methane (4,4'-H-3,5-HPM). Six of the 11 patients reacted to at least one of these compounds. All 6 patients reacted to 4,4'-H-3,3'-HPM and 5 reacted simultaneously to 4,4'-H-3-HPM. Four patients reacted to all 3 compounds tested. By patch testing with serial dilutions (ethanol 99.5%) of the three 4,4'-H-HPM, positive reactions were noted down to 0.5 X 10(-6)%, 1.0 X 10(-6)%, and 1.3 X 10(-3)% w/v for 4,4'-H-3-HPM, 4,4'-H-3,3'-HPM, and 4,4'-H-3,5-HPM, respectively.

This study compares the immune response of ten patients treated with polymerized grass (PG) immunotherapy with that of 12 patients treated with conventional monomer grass (MG) immunotherapy. The patients treated with PG immunotherapy received a mean cumulative dose of 44,840 protein nitrogen units (PNU), and those treated with MG immunotherapy received a mean cumulative dose of 46,083 PNU. Total antibody binding of perennial rye grass groups I, II, and III (RGGI, RGGII, and RGGIII) was measured in the serum of each individual. In addition, IgG titers to partially purified extracts of Bermuda, timothy, and orchard grass were also determined. A significant increase in the total antibody binding of RGGI, as well as an increase in the mean IgG titer to Bermuda, timothy, and orchard grass was demonstrated after treatment in the patients who had received PG immunotherapy. The mean total antibody binding of RGGII and RGGIII also increased in these patients, although not significantly. No significant difference in the mean total antibody binding of RGGI, RGGII, or RGGIII nor in the mean IgG titer to the three grass extracts was found in the sera of the two groups of patients. This study demonstrates a comparable immune response between immunotherapy with PG and MG, and retention of antigenic determinants during the polymerization process.

Mice which are genetic nonresponders to the random terpolymer of L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) not only fail to develop GAT-specific antibody responses when stimulated with soluble GAT either in vivo or in vitro, but develop GAT-specific T cells which suppress the GAT-specific plaque-forming cell response of normal nonresponder mice stimulated with GAT complexed to methylated bovine serum albumin (MBSA).Thus, both responder and nonresponder mice have T cells which recognize GAT. However, nonresponder mice can develop GAT-specific helper T cells if immunized with GAT bound to MBSA or to macrophages. The relevance of Ir gene-controlled responses is discussed.

The anti-tumour activity induced by glucans (lentinan, yeast cell walls, pseudonigeran, dextran, DEAE-dextran and dextran sulphate) and fructosans (levan and carboxymethyl-levan) was compared with the activity of C. parvum. The following effects on tumour systems in CBA mice were assayed: (a) adjuvant activity on the immune response against tumour-specific transplantation antigens (TSTA) with a methylcholanthrene-induced fibrosarcoma; (b) cytostatic activity of peritoneal macrophages against radiation-induced leukaemia cells; and (c) inhibition of tumour nodule formation in the lungs following i.v. injection of fibrosarcoma cells. All the polysaccharides induced cytostatic macrophages, but the dextrans and levans did so only after i.p. and not i.v. injection. Only lentinan, yeast cell walls and pseudonigeran were active in the lung-nodule inhibition test; and only lentinan and dextran sulphate showed slight adjuvant activity for TSTA. It is concluded that the anti-tumour activity induced by these polysaccharides is predominantly non-specific macrophage-mediated and much weaker than that found with C. parvum.

The authors studied the properties of two polymeres prepared from flagellin by various methods: precipitation with ammonium sulfate with the subsequent centrifugation or treatment with glutaric aldehyde in solution and subsequent gel-chromatography. Molecules of the former (P-I) and of the latter (P-II) were similar by shape, but the mol wt of P-I was greater. The preparations contained common H-antigen and were similar by serological activity. P-I had a very high "priming" activity for mice. P-II was highly immunogenic only within the narrow range of doses, was less immunogenic in comparison with P-II, and, in difference from P-I, induced formation of some amount of 7S-antibodies.

1310 patients were routinely patch tested with a paratertiary-butylphenol-formaldehyde resin (PTBP-F-R), a resol resin based on phenol and formaldehyde (P-F-R-2), and a mixture of these 2 resins. Approximately 2.5 times more patients with contact allergy to phenol-formaldehyde resins were diagnosed when routinely patch tested with P-F-R-2 in addition to PTBP-F-R. Although patch testing with a mixture of both resins was not as good as patch testing with the 2 resins separately, it was better than testing only with PTBP-F-R, since 1.6 times more patients with contact allergy to phenol-formaldehyde resins were still diagnosed. P-F-R-2 is therefore recommended for routine patch testing, preferably as a separate patch test but otherwise as a mixture with PTBP-F-R.

Thirteen patients with contact allergy to phenol-formaldehyde resins (P-F-R) were patch tested with 3-methylol phenol, 2,4-dimethylol phenol and 2,6-dimethylol phenol. Nine patients reacted to at least 1 compound, all giving positive test responses to 2,4-dimethylol phenol. Seven patients reacted simultaneously to 2,6-dimethylol phenol while only 1 patient reacted to 3-methylol phenol. Negative test responses were noted in 20 controls. Chemical investigation by high pressure liquid chromatography indicated that the compounds tested were pure and separable. The 3 reported sensitizers may, theoretically, be generated during the manufacture of P-F-R. 2,4-Dimethylol phenol and 2,6-dimethylol phenol have been demonstrated and there has been chromatographic evidence of 3-methylol phenol in the P-F-R used in the routine test series at the department.

Incorporated in oil-in-water emulsions, nonionic block polymer surfactants change the kinetics of generated antibody responses against pneumococcal hexasaccharide-protein conjugates: prolonged immunoglobulin M and immunoglobulin G responses are realized. Nonionic block polymer surfactants favor the immunogenicity of hexasaccharide-protein conjugates in young mice in such a way that a single injection yields long-lasting protection.

Polymers of different chemical structure were studied for their effect on the production of carcinogen-protein antigens (CPA) at early stages of benzidine-induced carcinogenesis in mice. Polyacrylic acid (out of the four polymers studied) was found to exert the most pronounced inhibitory effect on the CPA production mainly in the liver.

Extracellular polymeric material (EP) was isolated from culture supernatants of Candida albicans grown on carbon sources (50 mM-glucose, 500 mM-sucrose or 500 mM-galactose) known to promote yeast adhesion to different extents. Galactose-grown yeasts, which are the most adherent, produced more EP than sucrose-grown organisms, particularly after incubation for 5 d, while glucose-grown yeasts (the least adherent) gave the lowest yield. EP produced on all three carbon sources was of similar composition and contained carbohydrate (65 to 82%; mannose with some glucose), protein (7%), phosphorus (0.5%) and glucosamine (1.5%). Serological studies indicated that these EP preparations were immunologically identical but that galactose-grown yeasts had more antigenic determinants than sucrose-grown organisms while glucose-grown yeasts had the fewest determinants. Antigenic differences were apparent between EP preparations of some strains of C. albicans. Pretreatment of acrylic strips with EP to form a polymeric coating promoted yeast adhesion to the acrylic surface, but similar pretreatment of buccal epithelial cells with EP inhibited subsequent yeast adhesion. These results indicate that EP originates from the cell surface of C. albicans and that it contains the surface component(s), probably mannoprotein in nature, responsible for yeast adhesion.

The immune responses of the inbred guinea pig strains 2 and 13 have been determined against random terpolymers of L glutamic acid and L alanine and a third amino acid. Strain 2 guinea pigs responded against GAT10, GAT20(LLD), GAT10(NO2)15, GAT4, and GAL10. However, strain 13 guinea pigs responded only against GAT10. The explanation offered is that strain 2 guinea pigs, which have the Ir-GA gene, recognize the polymers via random GA determinants present in sufficient concentration in all of the above polymers. However, strain 13 guinea pigs recognize the GAT10 via the Ir-GT gene, and reduction in the concentration of tyrosyl residues below 10 mole % by various procedures alters the concentration of available random GT determinants necessary for interaction with the gene product of the Ir-GT gene.

Primary responses to the linear polymers of L-glutamic acid, L-tyrosine, and L-alanine are restricted to the IgG class of antibodies. The appearance of specific IgM antibodies against these antigens is dependent upon secondary immunization, in contrast to many classical antigenic systems. The presence of an IgM response was verified by a direct plaque-forming cell assay, the inhibition of direct plaques by an antiserum specific for mouse micron-chain, and the physical separation of IgM and IgG GAT-specific antibodies by gel filtration. Preimmunization of the appropriate nonresponder strain with GAT or GT inhibits both the secondary IgM and IgG responses to GAT-MBSA and GT-MBSA, respectively. The tolerance observed is due to the induction of suppressor cells as demonstrated by cell transfer experiments.

Pyran copolymer (NSC 46015) was evaluated with respect to its effect on the rejection of a murine leukemic allograft by BALB/c x DBA/2 F1 (CD2F1) mice. Significant prolongation of allograft survival with production of progressively growing lethal tumors was found following pyran administration. This phenomenon occurred at nontoxic doses of the drug and appeared to be closely related to the timing of pyran injection. Nonspecifically stimulated lymphocyte blast transformation by concanavalin A was not impaired by pyran when lymphocytes were exposed in vitro to the drug. The mechanism of tumor allograft enhancement remains obscure but may be related to allograft size at the time of pyran administration.

The biological response modifier maleic anhydride-divinyl ether (MVE-2) can activate natural killer (NK) cells and macrophages and can act as an immunoadjuvant for T and B cells. MVE-2 activates macrophages following intravenous or intraperitoneal injection in a compartmentalized manner, i.e., peritoneal macrophages (i.p. injection) or alveolar macrophages (i.v. injection). It activates NK cells in vivo but not in vitro, a dichotomy that may be secondary to interferon production. Splenic NK cell activity is not prolonged by the multiple injection of MVE-2; instead, it induces a state of NK cell hyporesponsiveness, which may limit its therapeutic efficiency. Therapeutic properties of MVE-2 are largely limited to nonspecific immunoprophylaxis, which may be associated with NK cell activation but which does not necessarily correlate with the level of splenic NK cell activation. Minimal therapeutic efficiency consisting of a slight prolongation in survival is observed in mice with preexistent disease treated with MVE-2. Prolonged survival is observed only in those animals placed on therapy soon after tumor cell challenge (experimental metastasis) and not in mice with established spontaneous metastasis. The need to manipulate the animal model (MVE-2 injection prior to or rapidly following tumor challenge) seems to predict that this agent is unlikely to be clinically useful against preexistent metastatic tumor burden, although some efficiency may be associated with local treatment into the pleural or peritoneal cavity.

The use of a biodegradable polymer for antigen delivery based on poly(CTTH-iminocarbonate) (IUPAC nomenclature: poly[oxyimidocarbonyloxy-p-phenylene [2-(hexyloxycarbonyl)ethylene]imino[2-[1-(benzyloxy)formamido]- 1-oxotrimethylene]-p-phenylene]) was investigated. This polymer was selected since its primary degradation product, N-benzyloxycarbonyl-L-tyrosyl-L-tyrosine hexyl ester (CTTH) was found to be as potent an adjuvant as complete Freund's adjuvant and muramyl dipeptide, when measuring the serum antibody response to bovine serum albumin (BSA) in male CD-1 mice over 56 weeks. BSA released from subcutaneously implanted polymeric antigen delivery devices made of poly(CTTH-iminocarbonate) resulted in significantly higher (P less than 0.006) anti-BSA antibody titers than devices made of poly(bisphenol A-iminocarbonate).

J chains purified from colostral IgA, polymeric (mainly dimeric) polyclonal IgA from serum, and monoclonal IgM appeared to be immunochemically identical in tests with antisera produced against the two latter antigens. None of the antisera precipitated with the native immunoglobulins, but J-chain antibodies became bound to all three types of Ig polymer. On a molar basis polymeric IgA was almost twice as efficient as IgM and 17 times more efficient than colostral IgA in antibody inhibition tests. The J-chain antibodies were able to link two or more IgA dimers and two or more IgM pentamers. After incubation in large antibody excess, the sedimentation properties of most IgM molecules were altered, whereas about 40% of the IgA dimers were unaffected by the treatment. THus, although the J chain on the whole was slightly more exposed in polymeric IgA than in IgM, it was more homogeneously available in the latter polymer. After denaturation in acid urea, the J chain became markedly more exposed in all three types of Ig polymer, and they were all precipitated by the most potent antiserum. Immunochemical quantitation of J chain released after reduction with 20mM dithiothreitol indicated that on the average there are about 2 mol of J chain per mol of polymeric IgA, and 3 to 4 mol per mol of IgM.

Spleen cells from adult CBA/H or CBA/N mice, or from neonatal CBA/H mice, were fractionated on thin layers of fluorescein (FLU)-gelatin to yield FLU-specific B lymphocytes. A single cell, or small numbers ranging from 1 to 10, were cultured in 10-microliter microcultures together with various antigens and mitogens. The results were compared with those of bulk culture or limiting dilution cultures supported by thymus filler cells. B cell growth and differentiation-promoting conditioned media (BGDA) were added to some cultures. The CBA/N results gave no support to the commonly used classification of T cell-independent (TI) antigens into TI-1 and TI-2 categories. A typical supposed TI-1 antigen, FLU-LPS, strongly stimulated normal adult single FLU-specific B cells to proliferate and form antibody, but virtually failed to trigger CBA/N B cells of comparable antigen-binding avidity. The same was true of LPS or LPS plus dextran sulfate acting as mitogens. The allegedly TI-2 antigen FLU-Ficoll, although still triggering comparatively poor responses, was actually marginally more active than FLU-LPS. FLU-Brucella abortus (FLU-BA) + BGDA gave the best results with single CBA/N B cells, but still induced only 1.27% of cells to develop into antibody-forming clones vs 12.2% with CBA/H cells. The results obtained with single neonatal B cells also lent no support to the distinction between TI-1 and TI-2. Both "TI-1" and "TI-2" stimuli caused adequate proliferation, one "TI-2" antigen stimulating 23.2% of the cells. None of the antigens caused good antibody formation, however, probably because multivalent antigens can deliver signals impeding the differentiation of immature B cells. It is therefore suggested that the classification of TI-1 antigens into two subcategories be abandoned, at least for the time being.

The offspring of low responder F344 female rats that were immunized with poly(Glu52Lys33Tyr15) aggregated with MeBSA prior to mating showed a higher antibody response to the polypeptide antigen than did the offspring of unimmunized females. Immunization of the mothers with unaggregated polypeptide, or with DNP-BGG, did not affect the antibody response of the offspring even when high doses of antigen were used. When the polypeptide used to immunize the mothers was given as an aggregate, some crossed the placenta to the fetus. The antigen was first detected in the placenta, blood and liver of the fetus at 15 days of gestational age. After birth, it was in the liver and spleen up to 6 weeks af age, and thereafter it was present only in the bone marrow.

Polystylene latex particles coated with serum albumin of various species, including non-primate serum albumin, were agglutinated by sera containing hepatitis B surface (HBs) antigen and hepatitis Be (HBe) antigen and by purified HBs antigen. Monomer and polymer albumin separated from human serum albumin preparations on latex particles were found to react with HBs antigen. Monomer from non-primate serum albumin preparations bound to latex particles was also found to have the ability to react with HBs antigen, but polymer of non-primate serum albumin did not. The mechanism of reaction between HBs antigen and the latex-bound serum albumin of various species is discussed.

The secretory component (SC) isolated from human milk was labeled with 2 mol of the fluorescent thiol reagent N-[7-(dimethylamino)-4-methylcoumarinyl]maleimide (DACM) per mol of SC through the reactive disulfide bond of SC. The binding of the labeled SC to polymeric immunoglobulins was examined by gel filtration by measuring the fluorescence of DACM at 478 nm. The labeled SC was bound to immunoglobulin M (IgM) and its (Fc)5 mu fragment and to dimeric immunoglobulin A (IgA). When the labeled SC was bound to IgM or the (Fc)5 mu fragment, the fluorescence of DACM increased about 30%. By use of this fluorescence change, quantitative studies were made on the equilibrium and kinetics of the reversible interactions of the labeled SC with two IgM proteins and their (Fc)5 mu fragments at pH 7.0 and 25 degrees C. All the IgM proteins and their (Fc)5 mu fragments had one binding site per mole of polymers. The affinity constant (6 X 10(8) M-1), the association rate constant (7 X 10(7) M-1 min-1), and the dissociation rate constant (0.1 min-1) of one IgM were different from those of the other IgM (1.7 X 10(9) M-1, 1.0 X 10(8) M-1 min-1, and 0.06 min-1, respectively). However, the values for the (Fc)5 mu fragments of the two proteins were the same (1.9 X 10(9) M-1, 1.1 X 10(8) M-1 min-1, and 0.06 min-1, respectively) and were very similar to those of the IgM with the higher affinity constant.(ABSTRACT TRUNCATED AT 250 WORDS)

A paratertiary-butyl phenol-formaldehyde resin is present in the routine patch test series of the International Contact Dermatitis Research Group. This resin was not sufficient to trace patients with contact allergy to resins based on phenol and formaldehyde (P-F-R). A particular resol resin (P-F-R-2) was demonstrated to be a good tracer of patients with P-F-R hypersensitivity. 2-methylol phenol (2-MP), 4-methylol phenol (4-MP) and 2,4,6-trimethylol phenol (2,4,6-MP) were, besides formaldehyde, known sensitizers in P-F-R. The sensitizing potentials were confirmed both in humans and in guinea pigs. Eleven new contact sensitizers in P-F-R were established. A consistently performed patch testing, including testing with serial dilutions of each sensitizer, facilitated comparisons and conclusions concerning (a) assessment of irritancy/allergenicity, (b) determination of highest acceptable contaminations, (c) evaluation of clinical relevance and (d) choice of substances for cross-reactivity testing. Six theoretically possible substances in P-F-R, 3-methylol phenol, 2,4-dimethylol phenol, 2,6-dimethylol phenol, 2,21-dihydroxydiphenyl methane (2,21-HPM), 4,41-dihydroxydiphenyl methane (4,41-HPM) and 2,41-dihydroxydiphenyl methane (2,41-HPM), were established as contact sensitizers and also demonstrated and identified in P-F-R-2. Five sensitizers were isolated from P-F-R-2 and identified to be 4,4-dihydroxy-3-(hydroxymethyl)-diphenyl methane (4,41-H-3-HPM), 4,41-dihydroxy-3,31-di-(hydroxymethyl)-diphenyl methane (4,41-H-3,31-HPM), 4,4-dihydroxy-3,5-di-(hydroxymethyl)-diphenyl methane (4,41-H-3,5-HPM), 2,41-dihydroxy-3,31-di-(hydroxymethyl)-diphenyl methane and 2,4-dihydroxy-31,51-di-(hydroxymethyl)-diphenyl methane. Nine sensitizers, 2-MP, 4-MP, 2,4,6-MP, 2,21-HPM, 4,41-HPM, 2,41-HPM, 4,41-H-3-HPM, 4,41-H-3,31-HPM and 4,41-H-3,5-HPM, were investigated using guinea pigs for sensitizations. The guinea pig maximization test was modified to increase standardization, enable a blind evaluation and also statistical comparisons between test animals and appropriate control animals. All 9 substances, except 4,41-HPM and 2,41-HPM, were established as contact sensitizers in guinea pigs. The 4,41-H-HPM were the most potent allergens. Phenol-formaldehyde resins and products based on P-F-R were investigated for the presence of 14 contact sensitizers. The concentrations varied and the highest concentrations were demonstrated for simple methylol phenols in P-F-R of the resol type.(ABSTRACT TRUNCATED AT 400 WORDS)

Gelatin-based plasma substitutes resemble denatured collagen in that sequence dependent antigenic sites of collagen may be preserved in these preparations and cause side reactions of such plasma substitutes. We have measured the binding of 125I-collagen in denatured conformation to antibodies and to cold-insoluble globulin (CIG) in sera of patients undergoing orthopedic surgery. About two thirds of the sera bound significant amounts of denatured collagen to immunoglobulins and to CIG. Yet, there was no correlation in individual patients or in groups of patients between the collagen binding capacity of sera and the occurrence of allergic reactions upon infusions of Haemaccel. Furthermore, antibodies to denatured collagen induced in animals did not cross-react with gelatin-based plasma substitutes. Thus, it is not likely that side reactions after infusion of gelatin-based plasma substitutes are caused by immunological reactions between antigens and performed antibodies.

The channels produced by complement in red cell membranes are heterogeneous, with diameters of approximately 0.5 to approximately 12.0 nm. We investigated the relationship of the components of the membrane attack complex, C5b through C9, to the functional transmembrane channels greater than 3 nm in diameter. Radiolabelled macromolecules were incorporated into resealed red cell membrane ghosts which were then treated with complement. A monoclonal antibody to a neoantigen in polymerized C9 inhibited macromolecule diffusion through the complement channels. There was also inhibition with polyclonal antisera to C9 but not with antisera to any of the other components of the membrane attack complex. The results demonstrate a functional correlation of the larger complement lesions with the previously described poly C9 tubular structures.

Specific multimer complexes of glycoproteins of enveloped viruses were prepared by treatment of suspensions of purified concentrated virus with a non-ionic detergent MECK mixed with 0.2% glycoside Quil A. Mixed complexes of glycoproteins with glycoside Quil A were formed upon removal of the detergent from the mixture of solubilized glycoproteins and glycoside Quil A. According to the results of electron microscopy, the formed complexes differed morphologically from the conventional micelles of glycoproteins and presented spherical reticular structures 20-30 nm in diameter. The immunogenic potency of the complexes was much higher than that of virus particles and micelles of purified glycoproteins and was comparable to the immunogenic potency of glycoproteins mixed with complete Freund adjuvant. The protective activity of the complex of protein G of rabies virus with glycoside Quil A was higher than that of the subunit rabies vaccine adsorbed on aluminium hydroxide.

This report characterizes the intra-H-2 crossover in the D2.GD mouse strain. Recombination occurred within the I region between the immune response (Ir) gene controlling immune responsiveness to ragweed extract (RE). Ir-RE, and the Ir gene governing responsiveness to the random linear terpolymers of glutamic acid and lysine with either tyrosine or pheylalanine (Ir-GLT and Ir-GLphi). The Ir-RE gene was tentatively located in the Ir-1A subregion and the Ir-GLT and Ir-GLphi gene(s) tentatively placed in the Ir-1B subregion. The importance of this recombinant strain to further studies on the fine structure of the I region is discussed.

Various molecular-weight maleic anhydride-divinyl ether copolymer polyanions were evaluated in six in vivo systems. Low- and high-molecular-weight MVE's were effective adjuvants with irradiated L1210 tumor cell vaccine. A high percentage of L1210-challenged survivors were refractory to a second challenge of tumor cells. Azimexon and Bacillus Calmette-Guérin were also effective adjuvants, but Bestatin was without adjuvant effect. All the MVE's demonstrated a marginal antitumor effect against the L1210 and LSTRA tumors. The MVW's, regardless of molecular weight differences, were effective in enhancing macrophage tumoricidal activity and retarding the development of M109 tumor growth in the lungs. Enhancement of delayed-type hypersensitivity by all six MVE's indicates their ability to stimulate T-cells.

The human IgA2-lambda myeloma protein Fel consists of covalent dimers and monomers which are partially self-associated. Affinity chromatography of this protein on staphylococcal protein A-Sepharose revealed that approximately 8% of the protein was retained and eluted by acid buffer. Although retained protein Fel was highly aggregated, in the presence of dissociating agents mostly monomeric form was found. Affinity rechromatography and electrophoresis of both fractions from affinity chromatography revealed that the retained fraction possessed substantially higher affinity for SpA than did the nonretained one. This could be due to the multivalency of protein Fel aggregates.

The multichain co-polymer poly(Tyr,Glu)-polyAla--polyLys, designated (T,G)-A--L, is normally thought to present only the poly(Tyr,Glu) sequences, designated (T,G), as antigenic determinants. Evidence is presented indicating that at least two different determinants in the (T,G)-A--L antigen are recognized by chickens. Studies in a partly inbred high-responder chicken strain reveal two major determinant systems: the (T,G) and the A--L in the (T,G)-A--L antigen. For one serum the antigenic determinant of the A--L backbone is shown to be a poly-D-alanine.

The adjuvant activities of four chemically similar, but physicochemically different nonionic surface-active agents called pluronic polyols F 68, L 31, L 101 and L 121 were studied. These four agents were tested in mice using an experimental model developed for studying the adjuvant activity of the cationic surface-active agent dimethyl dioctadecyl ammonium bromide (DDA). L 121 and DDA enhanced the primary antibody response to sheep red blood cells (SRBC) while F 68, L 31 and L 101 suppressed this response. The secondary humoral response to SRBC was enhanced by the polyol L 121 while the secondary response to dinitrophenylated bovine serum albumin (DNP22-BSA) was enhanced by both L 121 and L 101. DDA and the polyol L 101 were very effective adjuvants for induction of delayed-type hypersensitivity (DTH) to SRBC and DNP22-BSA after intracutaneous immunization of mice with a mixture of antigen and adjuvant. Since the four pluronic polyols were composed of identical chemical constituents, we proposed that difference in their activities as adjuvants were due to variation in their physicochemical properties. A correlation was found between a physicochemical parameter, the hydrophilelipophile balance (HLB), and the adjuvant activities of the pluronic polyols and several other types of nonionic surface-active agents. The agents which were strong adjuvants all had HLB values within a narrow range which classified them as spreading agents.

A covalent aggregate twice the size of insulin accounts for approximately 28% of total circulating insulin immunoreactivity in type I diabetic patients. These aggregates are probably covalent dimers of insulin and should contain unique epitopes distinct from the parent molecule. Therapeutic insulin contains a similar material and is the source of the circulating aggregate. Anti-aggregate antibodies were detected by binding-inhibition techniques in 9 of 29 long-term diabetic patients. These antibodies were directed against structures distinct from those of the parent molecule insulin monomer. All antibody-positive patients were men whose blood also contained antibodies to insulin monomer. We conclude that the blood of approximately 30% of insulin-using diabetic patients contains antibodies directed against epitopes unique to the insulin aggregates. Because insulin monomer and aggregates probably share a common primary amino acid sequence, the anti-aggregate antibodies are probably directed against conformational determinants. Further work is needed to determine whether such aggregates promote or accentuate the development of anti-insulin antibodies in certain genetically predisposed individuals.

Mice inoculated with both L1210 murine tumour vaccine and pyran copolymer were more resistant to L1210 than those inoculated with either of these agents alone. Rabbit anti-mouse thymocyte globulin and silica reduced the augmented resistance of these mice, suggesting the involvement of activated anti-tumour T cells and macrophages in the augmented resistance. We studied the activation of these two cells separately and examined the possible contribution of pyran copolymer-induced peritoneal cells to the augmented resistance to an inoculation of live tumour. Pyran copolymer-induced peritoneal cells endowed the tumour vaccine-primed mice, but not unprimed mice, with resistance to implanted L1210 and, among those peritoneal cell populations, macrophages but not T cells were responsible for this effect since the activity was associated with a cell population which was adherent to nylon wool columns, sensitive to silica and insensitive to anti-Thy 1.2 antibody plus complement. The pyran copolymer-induced peritoneal cells had very little antiproliferative activity when tested against L1210 in vitro and mice inoculated with these peritoneal cells did not survive a challenge of live L1210 cells much longer (less than 1 day) than L1210 inoculated control mice. Furthermore, the survival of L1210 vaccine-primed mice inoculated with one-tenth the amount of live L1210 (10(2)) was still much shorter than that of mice primed with L1210 vaccine plus pyran copolymer and challenged with ten times as many (10(3)) live L1210 cells. Therefore, direct tumoricidal activity was probably not a major factor in the in vivo immunological augmenting activity of the pyran copolymer-induced macrophages.

By combining oligopeptide sequences with synthetic polymeric chains, carriers of drug models can be prepared. The drug model (p-nitroaniline) is cleaved from the carrier in the lysosomal compartment of the cell. By changing the length and structure of the oligopeptide sequence, it is possible to regulate the rate of cleavage of drug model by individual thiol proteinases (cathepsins B, H and L) which are the most important as regards cleavage of the substrates studied. By connecting synthetic polymeric chains via oligopeptide bridges, it is possible to regulate the molecular weight as well as the biodegradability of the carrier molecule. Molecular weight also influences other biological properties, e.g., elimination from the organism, rate of pinocytic uptake and biological activity. Homopolymer of N-(2-hydroxypropyl)methacrylamide (HPMA) is nonimmunogenic in rats. Attachment of oligopeptide side chains gives rise to a macromolecule possessing immunogenic activity. The degree of antibody response depends on the detailed structure of the copolymer.

Three levels of humoral responsiveness (none, high or marginal) were noted following secondary immunization with random linear terpolymer of L-glutamic acid, L-lysine and L-leucine (GLleu). These immune responses were under dual H-2-linked Ir gene control. Gene complementation was observed in appropriate F1 offspring between two nonresponder strains and in selected intra-I region recombinants derived from crossing over between nonresponder parental haplotypes. One of the Ir-GLleu genes, beta, was localized within the I-A or I-B subregions. The other Ir-GLleu locus, termed alpha, was mapped in the I-C or S subregions. Differences in the strain distribution of immune responsiveness to GLleu and related terpolymer GLphe (which is also under complementing Ir gene control) are primarily attributable to differences among the Ir-GLleu and Ir-GLphe beta alleles. A correlation exists between the presence of the Ir-GLleu alpha(¿allele and the gene coding for specificity Ia.7.

Interest in the Ir genes of rheus monkeys stems from their phylogenetic relationship to man and the extensive data already available on the major histocompatibility complex of the monkey. At least two independent dominant H-linked Ir genes have been identified in the rhesus. These genes control the ability of monkeys to respond to the random linear copolymer of glutamyl alanine (GA), or the dinitrophenyl conjugate of glutamyl lysine (DNP-GL). These synthetic polymers can elicit weak delayed-type skin reactions and strong humoral responses in some monkeys. In a series of unrelated monkeys phenotyped for the serologically defined RhL-A specificities of both segregant series, there were no correlations between any RhL-A specificity and responder status to the GA or DNP-GL polymers. However, segregation analysis of 21 rhesus families sired by 3 fathers indicated the capacity of the offspring to form antibodies was associated with genes coded for in the RhL-A complex. In three monkeys, verified recombination within the RhL-A complex between the genes coding for the serologically defined determinants (SD loci) and the gene(s) controlling the lymphocyte-activating determinants (Lad loci) responsible for mixed lymphocyte reactivity was established. In two of these monkeys the immune response genes controlling the DNP-GL response segregated with the Lad genes, while in the third case the Ir-GL gene segregated with the SD loci, tentatively localizing the Ir-GL gene between the SD and Lad loci. In addition, we have shown that genetically distinct genes control responsiveness to DNP-GL and GA. These genes were separated by recombination, thus one monkey inherited the Lad, Ir-GL, and SD loci from one paternal haplotype and by crossing over inherited the gene controlling GA responsiveness from the other paternal haplotype. The fine structure mapping of the RhL-A gene complex is compared with the H-2 and HL-A gene complexes. Several striking similarities were noted.