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Some immunological parameters in rheumatoid arthritis from India.

The present study describes the immune status of 41 patients with rheumatoid arthritis from India. The results showed a high level of immunoglobulins, normal levels of C3, less proportion of seropositivity (54%) than that reported from the west and 26% having antinuclear antibody in their serum. T lymphocyte proportion in the blood was found to be low in acutely ill patients only. Mantoux and streptokinase-streptodornase skin tests for delayed hypersensitivity were positive only in about 50% of that of controls. However, all the patients could be sensitized to DNCB. For testing the humoral antibody response TAB vaccine was given. The levels of antibody produced against O and H antigens of "Salmonella typhi" were comparable in patients and controls. However, the level of IgM rose significantly in the patients in comparison to that of controls. The cause of this, at least partially, was found to be due to a very significant rise of rheumatoid factor after TAB vaccine in the patients. Also 3 out of 8 patients converted to a seropositive state after TAB vaccination. Controls did not show the development of rheumatoid factor.

Agglutinins↗

Differential ability of fixed antigen-presenting cells to stimulate nominal antigen-reactive and alloreactive T4 lymphocytes.

The capacity of paraformaldehyde-fixed human antigen-presenting cells (APC) to induce responses by autologous, freshly isolated peripheral blood T4 cells was examined and was compared with their ability to stimulate allogeneic T4 cell DNA synthesis. Fixation of glass-adherent cells (AC) with as little as 0.06% paraformaldehyde abolished leucine incorporation, whereas fixation with 0.75% paraformaldehyde caused death of greater than 98% of the AC. Control APC were able to take up and present the soluble antigens streptokinase-streptodornase (SK-SD), tetanus toxoid, or tuberculin-purified protein derivative to autologous Ia-depleted T4 cells. Fixation with greater than 0.06% paraformaldehyde eliminated such ability. When AC were incubated with antigen overnight and were then fixed, however, they were able to present nominal antigen to autologous T4 cells in a genetically restricted manner that was blocked by monoclonal antibodies directed against monomorphic determinants on class II major histocompatibility complex (MHC) molecules. Despite the ability to present nominal antigen, paraformaldehyde-fixed AC were unable to induce allogeneic T4 cell proliferation. Similar results were observed when non-T cells or spleen cells were used as stimulators. The inability of fixed APC to stimulate allogeneic T4 cell DNA synthesis was not reversed by increasing the number of fixed APC or by the addition of control AC autologous to the responding cells. Moreover, interleukins 1 and 2 either alone or in combination also failed to permit maximal T cell proliferation in response to fixed allogeneic APC. The differential effects of fixation on nominal antigen and alloantigen presentation could not be explained by the loss of membrane thymocyte stimulatory activity on fixed AC. These results indicate that antigen-bearing fixed APC are competent to stimulate proliferation by antigen-reactive T4 cells, but are deficient at inducing allogeneic T4 cell DNA synthesis. The differential sensitivity of these two Ia-restricted functions of APC to chemical denaturation (reductive methylation) by paraformaldehyde suggests that the allodeterminants and restriction elements for nominal antigen on MHC class II molecules can be functionally dissociated.

Antigen-Presenting Cells↗

Dissection of defective antigen presentation by interferon-gamma-treated fibroblasts.

The capacity of interferon-gamma (IFN-gamma)-treated HLA-DR expressing human dermal fibroblasts (FB) to function as antigen-presenting cells (APC) was examined. FB were cultured with 250 U/ml IFN-gamma for 4 days to induce HLA-DR expression. Peripheral blood monocytes (M phi), FB, or IFN-gamma-treated FB from the same donor were then cultured overnight with or without the recall antigen streptokinase streptodornase (SKSD), and their capacity to stimulate autologous T4 cell DNA synthesis was examined. SKSD-bearing M phi stimulated T4 cell proliferation, whereas antigen-bearing HLA-DR (+) FB did not. Even after fixation with paraformaldehyde to eliminate metabolic activity, SKSD-bearing M phi, but not FB, were able to function as APC. However, when HLA-DR (-) endothelial cell (EC) or autologous or HLA-D-mismatched M phi were added to the cultures, antigen-pulsed IFN-gamma-treated FB and M phi were comparably effective stimulators of autologous T4 cell DNA synthesis. Antigen recognition by the T4 cell was restricted by the class II major histocompatibility complex (MHC)-encoded gene products expressed by the IFN-gamma-treated FB and was unrelated to the class I or II MHC-encoded gene products expressed by the additional cell type. EC-promoted T4 cell DNA synthesis induced by antigen-bearing IFN-gamma-treated FB was inhibited by 60.3, a monoclonal antibody directed at an epitope common to LFA-1, CR3, and the p150,95 molecule. Inhibition caused by 60.3 was completely reversed by the addition of IL 2 to the cultures. Antigen presentation by IFN-gamma-treated FB was also enhanced somewhat by IL 1, IL 2, or monoclonal antibody directed at Tp44 (9.3). However, each of these additions alone promoted T cell proliferation less effectively than EC and resulted in responses that were smaller than those triggered by antigen-bearing M phi. The data suggest that IFN-gamma-treated FB take up and process antigen effectively, but lack an accessory cell property necessary for antigen-induced T4 cell IL 2 production and proliferation.

Antigen-Presenting Cells↗

Cell-mediated immunity in vitro in atopic dermatitis.

The function of T cells in atopic dermatitis was studied by leukocyte migration agarose and lymphocyte transformation tests. We found that phytohemagglutinin (PHA)- and PPD-induced release of leukocyte migration inhibition factor (LIF) from lymphocytes is significantly decreased (P less than .001) in patients with atopic dermatitis as compared with healthy controls. Blastogenic response of lymphocytes induced by PPD was also decreased in atopic patients as compared with controls (P less than .01). No differences were found in spontaneous blastogenesis or in blastogenic response of lymphocytes in vitro to PHA, concanavalin A (ConA), or streptokinase-streptodornase (SK-SD) between patients with atopic dermatitis and controls.

Adolescent↗

Immunology of atopic dermatitis.

Thirty-two adult patients with atopic dermatitis were studied at the Allergology Service of the "Hnos. Ameijeiras" Clinical Surgical Hospital. The diagnosis was established following the criteria of Hanifin and Lobitz. A detailed medical history was written for the patients; the study of some immunological parameters, such as the serum immunoglobulin quantification, delayed skin tests with a battery of antigens, and the spontaneous rosette-test, was also carried out. Almost all the patients showed serum IgE values above 150 UI, by means of the ELISA test modified by C.E.N.I.C. The mean values of the spontaneous rosette-test were low; this was more noticeable during the exacerbation period of the lesions. Candida sp, Mantoux and Streptokinase-Streptodornase antigens showed negative results in a high proportion of patients with atopic dermatitis, in relation with the control group. In atopic dermatitis, there are humoral disorders of immunity; this was demonstrated in our group by increased values of IgE and cellular disorders due to skin anergy, and to a low percentage of rosette forming cells; this does not allow to state that these phenomena have an active participation in the etiopathogenesis of this entity.

Adult↗

Functional abolition of monocyte HLA-DR by aldehyde treatment. A novel approach to studies of class II restriction elements in antigen presentation.

The effect of low concentration aldehyde treatment (0.0012 to 0.005%) on the expression of HLA-DR Ag by human monocytes was investigated. This treatment was shown to selectively abolish the expression of HLA-DR determinants defined by a monomorphic mAb (YE2.36) in a rosette assay. The expression of class I MHC Ag and Fc gamma R remained unaffected. As a result, the presentation of the recall Ag tetanus toxoid and streptokinase-streptodornase (SK-SD) to freshly isolated autologous T cells and T cell clones was completely inhibited. Increasing the concentration of aldehyde to 0.05% consistently produced partial restoration of Ag-presenting capacity. Low dose aldehyde treatment did not affect monocyte viability or membrane turnover. Thus, aldehyde-treated monocytes produced a second generation of HLA-DR and expression was almost completely restored to normal after 24 h of culture. The presentation of monocyte class II Ag as alloantigens was also inhibited by low dose aldehyde treatment but inhibition was much less marked when monocytes were aldehyde treated at 2 h rather than at 24 h of culture. This is consistent with the reexpression of HLA-DR which occurred readily in the first 24 h of culture and much less readily thereafter. Low dose aldehyde treatment did not affect Ag uptake and processing. However, monocytes which had been pulsed with Ag, aldehyde-treated to abolish HLA-DR, and then cultured to allow regeneration of HLA-DR could present Ag only when given a second Ag pulse, suggesting that once the association between microbial Ag and HLA-DR had formed the Ag was not then free to reassociate with novel HLA-DR. Low dose aldehyde treatment did not affect monocyte IL-1 production, neither did it inhibit the detection of HLA-DR by soluble mAb in FACS analysis. These results are consistent with the view that low dose aldehyde treatment disrupts the tertiary structure of human Ia molecules such that allostimulatory determinants and restriction elements for exogenous Ag are rendered inaccessible to T lymphocyte receptors and to cell-bound anti-DR mAb in the rosette assay, although DR determinants may remain accessible to soluble mAb.

Aldehydes↗

Blocking plasma factors of migration inhibition factor production in patients with cervical carcinoma. A possible role in vivo.

The blocking activity was studied of plasma from patients with cervical carcinoma in a model of migration inhibition factor (MIF) production by normal human leukocytes. At the same time, the patients' in vivo (intradermal reactivity) and in vitro (MIF assay) responses were studied with two common antigens (streptokinase-streptodornase and purified protein derivative). The plasma of 15 of 27 patients with carcinoma was capable of blocking the MIF test of human normal leukocytes. Twelve of 15 patients with blocking factor in their plasma had no or only a slight in vivo response, although they had a good in vitor response. Two patients had blocking factor without any in vivo or in vitro response. Six patients who had in vitro responses and slight or no in vivo responses had no plasma blocking activity. Three patients had neither in vivo nor in vitro responses and lacked blocking activity in their plasma.

Cell Migration Inhibition↗

In vitro proliferation of lymphocytes from human volunteers vaccinated with armadillo-derived, killed M. leprae.

A killed, armadillo-derived Mycobacterium leprae vaccine was examined for its ability to induce cell-mediated responsiveness in purified protein derivative (PPD)-positive volunteers residing in a nonendemic country using the lymphocyte transformation test (LTT). A marked increase in the proliferative responses to a M. leprae-soluble antigen preparation was observed in the two groups which were vaccinated with the highest doses of the vaccine, i.e., 1.5 X 10(8) and 5 X 10(8) bacilli. This increase was observed in both groups 3 months after vaccination, and persisted for the study period of 1 year. The in vitro proliferative responses to whole bacilli, of both armadillo and human origin, showed a similar but smaller increase 3 months after vaccination. Some enhancement of responses to cross-reactive antigens, such as PPD, and to unrelated antigens such as streptokinase-streptodornase, tetanus toxoid and diphtheria toxoid, was also observed. Thus, the LTT revealed that while the killed M. leprae vaccine induced a specific cell-mediated response to M. leprae, it was also responsible for a nonspecific immune-enhancement effect in healthy volunteers.

Adult↗

Abnormal cutaneous reactions in dermatitis herpetiformis.

In 94% of the patients with dermatitis herpetiformis, a locally applied ointment with an ester of nicotinic acid (Trafuril) induced an abnormal reaction with erythema, edema, papules, and often vesicles. The appearance of the reaction to Trafuril is similar to DH lesions. It differs markedly from the reactions to iodide where, as a rule, only one large blister results. The reaction is most pronounced 12-24 hours after application of Trafuril. The reactivity is strongest in the predilection areas for dermatitis herpetiformis. Strongly increased reactivity to Trafuril in DH is induced by pretreatment with streptokinase, streptokinase-streptodornase, or urokinase. Diminished reactivity to Trafuril is seen after pretreatment with injections of Compound 48/80, tranexamic acid, cromoglycate, or application of fluocinolone-acetonide, as well as after stroking the skin.

Adult↗

Lymphocyte response to phytohemagglutinin in atopic dermatitis.

Studies of patients with atopic dermatitis (AD) have demonstrated several clinical and laboratory indications of immunity defects, but with frequently contradictory results. We have recently shown that many patients with negative cutaneous delayed type hypersensitivity to candidin and streptokinase-streptodornase may have normal in vitro lymphocyte transformation to the same antigens. We hypothesized that this represents recovery of immunocompetent cells when they are isolated in vitro. This report describes phytohemagglutinin-induced transformation of lymphocytes immediately after isolation and after four days in culture (precultured). Responses of lymphocytes from patients with AD were initially subnormal, but increased to normal levels after the preculture period. Our results suggest that defective immune function in AD is not due to a permanent intrinsic lymphocyte defect, but is more likely due to factors associated with disease activity and severity.

Adolescent↗

Defective cell-mediated immunity in carcinoma of the prostate.

Cell-mediated immunity was assayed in 18 patients with a histologic diagnosis of carcinoma of the prostate and in 10 control patients with benign prostatic hypertrophy. In vivo cell-mediated response was assayed by intradermal skin testing to a battery of five delayed hypersensitivity antigens. In vitro cell-mediated immunity was studied by lymphocyte transformation using both heterologous and autologous "serums" and nonimmune rosette formation. Lymphocyte transformation studies were carried out using both specific antigens (Dermatophyton and streptokinase-streptodornase) and a nonspecific mitogen (phytohemagglutinin). Nonimmune rosette formation was performed under numerous temperature ranges. A defect in cell-mediated immunity was found in patients with carcinoma of the prostate. Skin tests demonstrated a significantly increased incidence of energy in cancer patients. Mitogen-induced lymphocyte transformation was significantly depressed when autologous serum from patients with carcinoma of the prostate was utilized in the procedure. It is concluded that defect in cell-mediated immunity does exist in patients with carcinoma of the prostate. This defect is expressed in vivo and in vitro and seems to be mediated by a serum factor.

Aged↗

Histoplasma meningitis with common variable hypogammaglobulinemia.

Histoplasma meningitis (HM) has been reported to occur primarily in association with disseminated histoplasmosis (DH). We report a case of histoplasma meningitis occurring in a patient with common variable hypogammaglobulinemia (CVH) in which no manifestations of DH were observed. L. L., a 66-year-old Caucasian male, clerical worker, developed occasional episodes of dizziness and tinnitus in mid-1971. During 1972, increasing frequency of these episodes and gradually progressive confusion were noted. In January 1973, vomiting, forther confusion, obnubilation, and a left central facial paresis developed and he was hospitalized. Physical examination revealed no pulmonary abnormalities, lymphadenopathy or hepatosplenomegaly. Over the ensuing 6-week evaluation, there was occasional fever to 38.5 degrees C. Chest roentgenogram was normal. Cerebral angiography suggested a mass in the left cerebellar hemisphere. EEG was diffusely slow. Multiple CSF examinations revealed: Glucose 7-18 mg/with a normal blood glucose, protein 109-256 mg/and cells 66-140 (95 + % mononuclear). Histoplasma capsulatum was cultured from CSF but not from sputum, urine, blood or bone marrow. Skin tests for PPD, histoplasmosis, coccidiodomycosis, blastomycosis, mumps, dinitrochlorobenzene and streptokinase-streptodornase were negative then and 6 months later. Histoplasma serum antibody was absent. Immunoglobulin analysis revealed IgG 430 mg %, IgA 46 mg %, and IgM 35 mg %, which with the history and skin test results suggested CVH. Treatment with 2.51 gm of amphotericin B given intravenously over a 3-month period resulted in complete reversal of all neurologic signs and clearing of the confusion. The remission has been maintained for two years. This case represents a primary infection of the CNS by histoplasma. The relationship between the HM and the CVH will be discussed.

Agammaglobulinemia↗

Modulation of the host response in human schistosomiasis. IV. Parasite antigen induces release of histamine that inhibits lymphocyte responsiveness in vitro.

Several mechanisms underlying the suppression of in vitro lymphocyte transformation responses to parasite antigens in human schistosomiasis have been previously described, but the role that immediate hypersensitivity reactions may have in regulating these lymphocyte transformation responses has been little explored. Using Hypaque-Ficoll-separated peripheral blood mononuclear cells (PBMC) from patients with schistosome infections, we found that histamine release could be demonstrated routinely in lymphocyte cultures challenged with adult worm, egg, or cercarial antigens. Release occurred within 1 hr of stimulation, and histamine persisted in the cultures for 6 days at levels of 10(-6) to 10(-7) M. That such concentrations were capable of suppressing LT responses in vitro was shown by the addition of exogenous histamine to modified PBMC culture systems from 10 normal individuals and eight patients with Schistosoma mansoni or Schistosoma mekongi infections. Responses to phytohemagglutinin, streptokinase-streptodornase, and tetanus were equivalently suppressed in both groups (50.8 +/- 6% in normals and 55.9 +/- 6.2% in patients), and the doses required for maximal suppression were similar. Passage of PBMC from infected patients over nylon wool, in addition to removing adherent suppressor cells, also markedly reduced the number of histamine-containing basophils (74 +/- 4.5% removed). The enhanced responsiveness to parasite antigen by PBMC depleted by nylon wool passage was abrogated by the addition of exogenous histamine to the cultures. These results indicate that in routine PBMC cultures 'nonspecific' lymphocyte suppression by histamine liberated from basophils in an antigen-specific fashion may help to account for the specific suppression of lymphocyte responses to parasite antigens so characteristic of patients with schistosome and other helminth infections.

Antigens↗

Antigen-specific lymphocyte transformation in patients with recent yersiniosis.

Lymphocyte transformation in patients with recent yersiniosis was studied. A micromethod using washed blood cells and Yersinia enterocolitica antigen was employed. The washed blood cells were incubated in the presence of various dilutions of heat-treated whole bacteria; these proved as antigen superior to gentamicin- or formalin-treated bacteria. Patients with recent yersiniosis had a significantly higher response against Yersinia antigen as compared to 20 healthy controls, who had either no response or a low response. No difference could be observed in responses against PPD or streptokinase-streptodornase, or in the mitogen responses between these two groups. A marked cross-reaction was observed between Yersinia and Escherichia coli antigen. The results show that patients with recent yersiniosis develop lymphocyte transformation response against Yersinia. Lymphocyte transformation test can be used in the study of host responses against infecting Yersinia in patients with different clinical pictures of yersiniosis.

Antigens, Bacterial↗

Subcorneal pustular dermatosis--an unusual course.

In a patient with subcorneal pustular dermatosis (SPD) the appearance of new pustular lesions characteristic of the disease was triggered by two episodes of drug eruption (induced by dapsone and quinidine sulphate respectively) and by the intra-dermal injection of the recall antigens Candida and streptokinase-streptodornase, performed for evaluation of delayed hypersensitivity. Both episodes of drug eruption were probably the result of an immediate-type hypersensitivity reaction towards the offending drugs, as indicated by positive mast cell degranulation tests.

Aged↗

Autologous and allogeneic suppressor lymphocyte inhibition of human erythroid colony forming unit proliferation.

Certain in vitro and in vivo animal studies have supported the concept that lymphocyte-derived microenvironmental factors are important in erythroid proliferation. Considerable controversy has developed regarding the applicability of these concepts to human erythroid proliferation. We, therefore, evaluated the role of lymphocytes on human erythroid colony forming unit (CFUE) proliferation in the plasma clot system. Bone marrow (BM) was obtained from normal donors and cocultured with the following cell populations: a) cultured (6 day) lymphocytes autologous or allogeneic to BM; b) cultured lymphocytes stimulated with conconavalin A (Con A), allogeneic lymphocytes or streptokinase streptodornase (SKSD). In general, unstimulated cultured lymphocytes enhanced CFUE proliferation. In contrast, lymphocytes stimulated with Con A. SKSD, or allogeneic lymphocytes suppressed lymphocyte proliferation. The suppressor cell was concentrated in the T-cell fraction obtained by sheep red blood cell rosetting followed by ficoll-Hypaque centrifugation. Both allogeneic and autologous stimulated T-cells suppressed CFUE. Moreover, supernatants from stimulated lymphocyte cultures suppressed CFUE proliferation although the cell of origin and characteristics of the suppressive factors have not been defined. These data support the concept that lymphocytes may play an important role in modulating the human BM erythroid microenvironment.

Bone Marrow Cells↗

Abnormalities of lymphocyte transformations in women with condylomata acuminata.

The cellular immunity of 16 women with recalcitrant condylomata acuminata was investigated with an in vitro assay of antigen-induced lymphocyte transformations. Results were compared to those of 24 age-matched controls. Lymphocyte transformation responses (in counts per minute) for women with condylomata were much lower than controls for phytohemagglutinin-P, 32,285 and 60,015, (P less than .001); for concanavallin A, 28,664 and 58,605, (P less than .001); and for pokeweed mitogen, 34,941 and 73,394 (P less than .0001). No significant differences in lymphocyte transformations were noted between the 2 groups to Candida or streptokinase-streptodornase antigens. Immunosuppressive diseases, recurrent infections, and intraepithelial neoplasms of the genital tract were more frequent in the group of women with condylomata, and this seems to clinically substantiate the depressions noted in their in vitro cellular immunity. A refractory course of condyloma acuminatum in some women may be a reflection of an underlying state of immunosuppression.

Adolescent↗

Suppressor T cell numbers and function in atopic patients with high serum IgE levels.

Suppressor T cell function and the sensitivity of lymphocyte transformation to histamine has been studied in 9 atopic patients with high serum IgE levels and in 14 controls. The effect of concanavalin A (Con A) induced suppressor T cells on the proliferative response of fresh lymphocytes to mitogens and antigens was measured; in atopic subjects the median suppression to 5.0 micrograms/ml Con A was 80% to 1.0 microgram/ml Phytohaemagglutinin (PHA) was 73%, to Dermatophagoides pteronyssinus extract was 45% and to streptokinase--streptodornase was 24%. Indomethacin increased lymphocyte proliferation to mitogens, but to a variable degree, in both groups. Histamine suppressed lymphocyte transformation to PHA and Con A (median suppression in normal subjects 38 and 46%, and in atopics 51 and 60%) and to D. Pteronyssinus extract. There was no significant difference between normal subjects and atopics in any of these functional assays. The relative and absolute numbers of total T cells, helper T cells and suppressor T cells measured by monoclonal antibodies and the helper to suppressor T cell ratio were normal in the atopic group. These results show that the activity of Con A induced suppressor T cells and the effect of histamine and indomethacin on lymphocyte proliferation is normal in highly atopic subjects. No suppressor T cell defect has been identified using these assays.

Adolescent↗