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A 60-year-old man with dermatomyositis was admitted to our hospital because of dyspnea and hypertension. He had high fever and convulsive seizures after admission. Laboratory examinations showed hemolytic anemia, thrombocytopenia, and renal failure. A clinical diagnosis of thrombotic thrombocytopenic purpura (TTP) was made. He failed to respond to plasma exchange therapy, pulse therapy with methylprednisolone, high-dose gamma-globulin therapy, and antiplatelet therapies with ticlopidine, dipyridamole and a prostacyclin analog of beraprost sodium. He died on his 17th day in hospital. Autopsy examination revealed widespread microthrombi in his kidneys, lungs, spleen, and intestine. Only seven cases of dermatomyositis or polymyositis complicated by TTP have been cited in the literature. TTP was fatal in 6 of these 7 cases. Early diagnosis and prompt treatment may improve the outcome of TTP patients with dermatomyositis. Dermatologists should keep in mind that TTP occasionally arises as a serious complication of dermatomyositis.

Conjunctival tissue of 12 guinea pigs (6 experimental animals, 6 controls) was sensitized with IgG1 (PCA titer 1:2,000) antisera to dinitrophenol carrier and challenged topically with hapten. Periorbital swelling, conjunctival edema and conjunctival redness were evaluated clinically at 0.5 h and hourly for 12 h, and again at 24 and 48 h. Tears were collected at each time point for cytology. Results showed an early-phase reaction (EPR) peaking at 0.5 h and a late-phase reaction (LPR) peaking at 5-7 h. Periorbital swelling was minimal 4 h after appearance of the EPR. Conjunctival edema and redness increased in both the early and late phases, but without two distinct peaks. However, individual animals in both groups did show biphasic reactions. Tear cytology showed an abnormal increase in neutrophils and eosinophils in the later time periods and was a significant way to detect ocular anaphylaxis. In conclusion, if the presence of two significant peaks of clinical inflammation after one antigen challenge is taken as the narrowest definition of a late phase in anaphylaxis, our results show an ocular LPR occurring in our animal model.

An active model of ocular anaphylaxis was developed in guinea pigs to evaluate the histopathology of the early (EPR) and late (LPR) phase reaction, focusing on the role of mast cells. Five groups (n = 6) of animals were actively immunized by first injecting into each of the axillary and inguinal lymph node areas, 0.25 ml of an emulsion containing 1 mg dinitrophenyl bovine gamma-globulin (DNP-BCG) with 0.5 ml complete Freund's adjuvant. After two weeks, an intramuscular injection of 0.5 ml of an emulsion containing 1 mg DNP-BGG with 0.5 ml of incomplete adjuvant was administered. One month after the first injection, animals were sacrificed after topical ocular challenge with 10 microliters of 1 mg/ml divalent hapten, di-DNP-lysine, in one eye and phosphate buffered saline (PBS) in the fellow eye as control. Clinical reactions were graded over time and histology evaluated at the endpoint (time 0, 0.5, 3, 9, and 24 h). Results showed that all animals clearly developed both an EPR and an LPR, as either a biphasic, multiphasic or prolonged clinical response. A small percentage of mast cells were degranulated at baseline, whereas, at 0.5 h, 95% of mast cells were degranulated in the eyes treated with specific hapten and 25% in the control eyes treated with PBS. At 3 h, 84% of the mast cells were degranulated. This value rose to 89% at 9 h, and remained unchanged at 24 h.(ABSTRACT TRUNCATED AT 250 WORDS)

The present study investigates the question of capillary recruitment and reserve capillaries in the brains of awake rats. Perfused capillaries were marked by intravenous globulin-coupled fluorescein isothiocyanate (FITC) and cerebral blood flow was measured autoradiographically. During hypercapnia, the density of perfused capillaries was unchanged compared to normocapnia, although blood flow was markedly increased. This shows the lack of capillary recruitment in the brain during the high flow that occurs during hypercapnia. In additional studies using fluorescent staining both of morphologically existing and of perfused capillaries, perfusion of all capillaries during normal, normocapnic conditions was found. These experiments show the lack of any capillary reserve in the rat brain.

A model of topically induced ocular anaphylaxis in guinea pigs was developed. A guinea pig anti-dinitrophenyl (DNP) bovine gamma-globulin antiserum rich in IgG1 antibodies was injected subconjunctivally; various serum dilutions and latent periods were tested. The anaphylactic response was elicited with di-DNP-lysine applied topically. The response was characterized by rating 11 ocular signs. Although the 4-hour latent period is considered optimal for passive cutaneous anaphylaxis, a 14-hour latent period yielded the strongest ocular reaction and appeared to the most appropriate time for clinical assessment. Both an early-phase reaction (peaking in the 21st hour) and a late-phase reaction (peaking at 6-10 h) were observed. The two signs of the early-phase reaction that recurred in the late-phase reaction were periorbital swelling and lid redness. The anaphylactic reaction displayed different patterns in different animals: protracted, biphasic and multiphasic.

The properties of macrophage migration inhibition factor (MIF) and mitogenic factor (MF) were compared using culture supernatants of antigen-stimulated lymph node cells from inbred guinea pigs. Gel filtration on Sephadex G-100 indicated molecular weights of about 60,000 and 25,000 for MIF and MF, respectively. The lymphokines also differed with respect to heat sensitivity, MIF being largely inactivated by 60 degrees C for 20 min, whereas MF was unaffected by this treatment. The time course of production, antigen specificity of induction, and susceptibility to neuraminidase were also examined.

The effects of lysozyme and sodium diethyl-dithiocarbamate (DTC) were studied on antibody synthesis induced by immunization with a hapten-carrier complex. The immunomodulating effects of lysozyme could be partially reversed by DTC, a sulphur derivative which has been previously shown to modulate the immune response, probably at the macrophage level. The mechanisms whereby lysozyme and DTC modulated the immune response are discussed.

The "single-stage" mixed antiglobulin reaction (MAR) was carried out with rabbit thymocytes. This test involved treating the cells with either sheep or goat anti-rabbit globulin sera, and subsequently reacting them with indicator erythrocytes coated with rabbit immunoglobulin (Ig) so as to form rosettes. An unexpectedly high number (up to 38%) of thymocytes reacted, although the rosettes were weaker than those given by peripheral B lymphocytes. When blood and lymph node lymphocytes or thymus cells which had already been treated with sheep anti-rabbit globulin serum were subsequently exposed to rabbit anti-sheep Ig serum and then rosetted with indicator cells coated with ox Ig (cross-reacts with sheep Ig) almost 100% reaction was obtained in each of the cell suspensions. This was designated the "two-stage" MAR. The anomalous results, both in the one-stage and two-stage MAR, were abolished by pepsin-treating the sheep anti-rabbit globulin serum; thus indicating that sheep Ig is adsorbed non-specifically via the Fc part of the molecules to the surface of rabbit thymocytes and peripheral T lymphocytes.

Antibodies to bovine gamma-globulin (anti-BGG antibodies) were detectable by a radio-immunoassay in 70% of healthy blood donors but, generally, the titres were low. Significantly increased concentrations of anti-BGG antibodies were found in patients lacking IgA but not in patients with allergic disorders. The anti-BGG antibodies were shown to give rise to falsely high IgE values in the radio-immunosorbent test for IgE determination (RIST) when a sheep anti-IgE antiserum was used. Furthermore, falsely positive results can sometimes be caused by such antibodies in the determination of cow-dander- or cow's-milk-specific IgE by the radio-allergosorbent test (RAST). When a rabbit anti-IgE antiserum was used instead of the sheep anti-IgE, normal IgE levels and negative RAST results were obtained. The difference is explained by the higher degree of cross-reactivity between the anti-BGG antibodies and sheep alpha-globulin than between anti-BGG antibodies and rabbit alpha-globulin.

The spontaneous fixation of penicillin G on guinea pig serum protein was studied. Serum albumin and Ig were shown to fix firmly over 95% of penicillin. Therefore, guinea pigs were immunized with penicillin alone or penicillin-autologous carrier protein conjugates. The production of hemagglutinating antibody, of MIF and the in vitro stimulation of lymphocytes were analyzed as immunological parameters. The results showed that in all cases the strongest immunological response was obtained with animals immunized with the protein-hapten conjugates, whereas no response could be obtained with the carrier protein alone. Finally it was shown that the conjugates significantly stimulated purified T cell population. These results suggest that the antigenic recognition of penicillin involves the participation of lymphocytes.

An immunologically unresponsive state induced in experimental animals can be modulated by a number of procedures. Such modulation can result in the termination of the unresponsive state if there exists a normal complement of immunocompetent B cells in the presence of unresponsive T cells. It is possible to bypass the T cells and activate the B cells to respond to otherwise tolerated antigens. The bypass results from supplying a second (nonspecific) signal to the B cells, which, in the presence of antigen (specific signal), permits the B cell to differentiate. The role of the T and B cells and their activation and/or inactivation in the fate of both leukemic cells and oncogenic viruses is discussed.

Footpad reactions elicited in DHS mice using human gamma-globulin (HGG)-coated polystyrene latex particles and soluble HGG (sHGG) were compared. Both the magnitude and the persistence of DHS lesions produced by HGG-latex were considerably greater; at 24, 48 and 72 h after challenge, the level of reactivity induced by HGG-latex was 24, 41 and 71% higher, respectively, than those elicited with sHGG. Early nonspecific swelling following injection of HGG-latex in footpads of normal mice was negligible by 24 h, whereas Arthus-responsive animals did not return to control levels until 48 h. The possible advantages and limitations of using particle-associated protein to elicit DHS reactions are discussed.

Six normal rabbits were injected intravitreally with bovine gamma-globulin (BGG). Quantitation of immunoglobulins A, M and G, in the aqueous and in the serum, was performed by the single radial immunodiffusion following the method described by MANCINI et. al. Concentration of anti-BGG antibodies was determined by the reversed Mancini method. The antibody quotient was calculated; it reached to 8 and 5.96 in the aqueous of the injected eyes 10 days after intravitreal injection, 22.69 and 24 days, 22.52 after 28 days and 14.93 after 31 days.

Rabbit anti-chicken gamma-globulin was labeled with 125I and then incubated with cells from the bursa, thymus, spleen, and bone marrow of 4- and 8-week old birds. The same procedure was carried out on 11-week-old agammaglobulinemic chickens. Autoradiography revealed that the majority of large, medium, and small bursal lymphocytes bind the antibodies while labeled lymphocytes of each type in the spleen and thymus never exceeded 11 or 4 percent, respectively. Labeled medium and small lymphocytes in the bone marrow increased from 4.2 and 1.7%, respectively, at 4 weeks of age, to 9.5 and 8.8%, respectively, at 8 weeks of age. Labeled lymphocytes of all sizes were completely absent in all tissues of agammaglobulinemic chicks, including the marrow. Therefore, the increase in frequency of labeled lymphocytes in the bone marrow with age may be a result of recruitment of cells from the bursa of Fabricius. The majority of lymphocytes in the bone marrow do not label. Therefore, lymphocytes from the bone marrow may be T cells, subsets of B cells, or neither T or B cells.

Subcutaneous injections of a mixture of dinitrophenylated ovalbumin (DNP3-OA) and dextran sulfate into Swiss-Webster mice elicited short-lived primary and long-lasting secondary IgE antibody responses to both DNP and OA. Histamine was released on in vitro challenge with antigen (OA or DNP22-BSA) of washed peritoneal mast cells (PMC) obtained from mice during a primary or a secondary IgE response. Administration of an intravenous injection of a tolerogenic conjugate of DNP8-mouse gamma-globulin, either prior to immunizationor during an ongoing IgE response, resulted in almost complete disappearance of circulating anti-DNP IgE antibody and in a very marked decrease in histamine release from PMC on challenge with DNP22-BSA. However, the IgE response to OA of these mice and the histamine release from their PMC on challenge with OA were not affected. Moreover, the PMC of mice, which had been tolerized to DNP, could be passively sensitized with serum containing DNP-specific IgE antibody for the release of histamine on DNP22-BSA challenge. The most significant finding of this study is the observation that the time course for the loss of reactivity of PMC to DNP22-BSA, after administration of the tolerogen during an ongoing secondary response, paralleled the decrease in circulating anti-GNP IgE antibody.

SJL mice became resistant to tolerance induction between the 2nd and 6th week of life. The effect on tolerance induction of radioresistant accessory (A) cells does not depend on the age of the donor, i.e. is the same whether the A-cell donor is 3 or 12 weeks old. In the thymectomized recipient, reconstituted with spleen cells of 12-week-old donors, resistance to tolerance is greater than in the corresponding intact animal. Resistance to tolerance is due to a change in the T cells, occurs at a different rate in the T cells of thymus and spleen, and my be due to change in the relative proportion of different T-cell subpopulations.

Delayed type skin reactions in guinea pigs have been assessed by measuring three parameters: increase in skin thickness, diameter of erythema and intensity of erythema. Some groups of animals were immunized with different protein antigens in Freund's complete adjuvant with or without cyclophosphamide (CY) pretreatment; others received a single high dose of antigen intravenously at the time of immunization. The results emphasize the importance of measuring all three parameters for several days after skin testing. The intensity or erythema was found to be an especially useful parameter for assessing CY-induced modification of delayed hypersensitivity (DH) reactions. The increase in skin thickness, which can be measured objectively, was also valuable as both immediate and DH reactions are characterized by induration. The diameter of erythema could only be measured accurately for a short time after skin testing. Furthermore, the effects of intravenous antigen and CY pretreatment were not reflected by that parameter.

The modulating effects of passive antibodies on both delayed hypersensitivity to the carrier and antibody synthesis to carrier and hapten determinants were studied in guinea pigs. Animals were injected with antibodies directed against either the carrier or the hapten prior to immunization with the hapten-carrier conjugate in Freund's complete adjuvant. Anti-hapten antibodies have been shown to have an enhancing effect on delayed hypersensitivity to the carrier and a suppressive effect on antibody synthesis to the hapten. In this experiment, anti-carrier anti-bodies seemed to have had no effect on delayed hypersensitivity to the carrier and on antibody synthesis to the hapten.

Relationships between Normotest, prothrombin time, albumin, gamma-globulin, GPT, and the hepatic fractional clearance of 198Au-colloids as a measure of effective hepatic blood flow were studied by correlation analysis, in 50 patients with liver diseases, including chronic hepatitis or cirrhosis. Simple correlation coefficients were highly significant between Normotest, prothrombin time, albumin, and the hepatic fractional clearance; however, not between GPT or gamma-globulin and the other parameters. Further correlation analysis revealed a highly significant partial correlation coefficient between Normotest and albumin and the hepatic clearance; however, not between Normotest and albumin. As effective hepatic blood flow is considered to be proportional to the liver parenchymal volume, the above results indicate that vitamin-K-dependent clotting factors, as well as albumin, reflect the liver cell mass.

The immunologic effects of ampicillin polymers were studied in mice and rabbits. Polymerized ampicillin--in contrast to monomers--had a suppressive effect on the antibody response to penicilloylated proteins. Unfractionated ampicillin polymer, and high molecular weight fractions were found to have an immunogenic effect in mice, when tested by a haemolytic plaque assay. The immunogenic effect of variously sized polymers did not correspond to their antigenic activity as reflected by passive cutaneous anaphylaxis experiments.

An attempt to provide an overview of intrinsic problems encountered in the definition of the immunochemical parameters involved in the allergic response in terms of the appropriate in vivo and in vitro manifestations. The primary difficulty stems from the complexity of the chemical composition of most extracts of allergenic materials and from the lack of simple laboratory methods for establishing the allergenic profile of each patient as a function of the different components of a given extract. The principles of inhibition of the Prausnitz-Küstner passive transfer test and of the radioallergosorbent technique will be described; from the results one may deduce the minimum number of distinct allergenic components and of the corresponding reaginic antibodies of IgE type. The possible use of monovalent, hapten-like, low molecular weight constituents of allergens for blocking the triggering of the release of vasoactive amines from mast cells and basophils of allergic patients will be discussed. In addition of IgE antibodies, the spectrum of antibodies produced by the allergic patient, both before and after hyposensitization, involves immunoglobulins of the IgG, IgM and IgA classes, which can be measured by radioimmunoelectrophoretic and -immunodiffusion techniques; IgM antibodies can be readily differentiated from IgG antibodies by the hemagglutination technique, or by adaptation of reverse immunosorbent procedures. Similarly, the heterogeneity in relative affinities of the different antibodies for a given allergenic fraction may be established by the use of immunosorbents. The effectiveness of hyposensitization therapy could be thus correlated with objectively measurable immunological parameters. Possible mechanisms responsible for the effectiveness of this treatment will be discussed in the light of recent experiments in animal systems, suggesting the mandatory cooperation between T and B cells for the induction of the reaginic response.