Use of immunoenzyme analysis in the rapid diagnosis and study of the characteristics of circulating O-antigen in biological fluids of patients with acute dysentery Sonne

The data on the clinical approval of the original enzyme immunoassay system for the determination of somatic O-antigen in the blood serum and urine of patients with acute Sonne dysentery are presented. The level of the antigen determined in the biological fluids of patients has been shown to depend on the severity of the disease. Different types of dynamic curves, reflecting the level of O-antigen in the biological fluids of patients with acute Sonne dysentery and characteristic of different clinical forms of the disease, have been established.     

Indicators of spontaneous and stimulated nitroblue tetrazole test of polymorphous-nuclear leucocytes in acute dysentery patients

140 healthy individuals and 93 sick with acute dysentery were subjected to an examination by spontaneous and by bacterial preparations stimulated reaction with nitroblue tetrazole (NBT test). Indicators in healthy persons were normal in the spontaneous, and increased in the NBT test, stimulated by bacterial preparations. Indicators of the spontaneous NBT test in patients with acute dysentery were raised with a maximum in the period of early convalescence. Stimulation by a live shigella culture--the dysentery vaccine--revealed by means of Sonne diagnostic high, and when endotoxin from Serratia marcescens and dysenterin was used as an inductor, mild indicators of NBT test activity. When a polyvalent agglutinating dysentery serum was used as a stimulator, the activity increased considerably, and a simultaneous use of serum and vaccine had an inhibiting effect on the indicators of the stimulated NBT test. The obtained results testify the sufficient high reserve possibilities of leucocytes towards complete phagocytosis and the efficiency of the NBT test, stimulated by bacterial preparations for the study of functional and metabolic activity of leucocytes in the process of acute bacterial dysentery.     

Immunological diagnosis of Mycoplasma pneumonias]

A complex of immunological cell tests with M. pneumoniae antigen (the lymphocyte blast-cell transformation test, the allergic neutrophil alteration test) was carried out in order to establish the correlation between the results of positive seroconversion and the sepcific immunological reactivity of lymphoid cells in pneumonia patients. Mycoplasmic cutireactive allergen, when used for the accelerated diagnosis of mycoplasmic pneumonia in humans, was shown to be specific and safe. Cuti-allergic tests with mycoplasmic allergen allowed to diagnose mycoplasmic pneumonia at early stages (beginning from days 5--7), which ensures the possibility of indicating etiotropic treatment to patients in due time.     

Dynamics of bacterial polysensitization in acute dysentery in children depending on the species of the causative agent

Studies on polysensitization, evaluated by the neutrophil damage index, have been made with the use of dysenterin and the allergens of hemolytic staphylococci, enterococci and Escherichia coli. The studies have revealed that both specific and nonspecific sensitization to hemolytic staphylococci and enterococci is higher in dysentery caused by Shigella flexneri than in dysentery caused by S. sonnei and has the tendency to increase in the course of the disease.     

Several allergic reactions under conditions of functioning of an inhibitor of antibody activity in the body]

Allergization under conditions of functioning an inhibitory factor in the organism was studied by the reaction of leukocytolysis with Tsuverkalov's dysenterin and in the reproduction of hypersensitivity of the immediate type on the Schultz-Dale's model. It was shown that in functioning an inhibitory factor in the organism the degree of leukocytolysis reaction with the dysentery allergen decreased in the immunized animals in parallel with the fall in the serum antibody activity. A depression of the activity of macromolecular sensitizing antibodies was also observed.     

Lymphotoxin production by human lymphocytes. III. The action of adenovirus antigen]

In lymphocyte cultures (LC) from 5 immune donors blast transformation occurred as a result of the stimulting action of adenovirus antigen; the supernatants obtained from these cultures produced a cytotoxic effect on L cells, thus suggesting the presence of lymphotoxin (LT). No blast transformation and no LT production were detected in nonstimulated LC used for control, as well as in LC from 9 nonimmune donors. LT production was shown to depend on the time elapsed after recovery from acute adenovirus infection. LT production was found to be most intensive during the first 2 months after recovery.     

Immunologic activity of human lymph cells in the lymphocyte blast transformation reaction

The immunologic activity of lymph cells obtained by curative draining of the chest lymph drainage was studied in 14 patients with subhepatogenous jaundice of different origin. The lymphocyte blast transformation reaction showed that lymphocytes of the lymph can form blasts under the effect of phytohemagglutinin, antilymphocyte gamma-globulin and some bacterial antigens. The reaction of the highest intensity was recorded in lymphocyte cultures of the lymph collected for this purpose on the 2--3d day from the commencement of lymph drainage. Lymphorrhea produces a decrease in the capacity of stimulated lymph cells for blastformation. During lymph drainage no blastformation takes place in stimulated blood cell cultures of the patients. Therefore, the fundamental part of antigen sensitive lymphocytes is stored in the lymphoid tissue.     

Preliminary lymphocyte activation by a mitogen as a condition for demonstrating the ability of the Fab-fragment of normal IgG to inhibit lymphocyte transformation

Peculiarities attending inhibition of the PHA-induced blast-cell transformation of human lymphocytes by F(ab')2 fragment of rabbit IgG were studied. It was shown that the fragment did not affect the intensity of blast-cell transformation if the lymphocytes were preliminarily incubated with the fragment for 24 h at 37 degrees or 4 degrees C and then transferred to the fresh medium containing PHA. However, if the fragment was added to the cells 24 or 48 h following PHA it produced a significant inhibition of the blast-cell transformation. These data may indicate that F(ab')2 fragment interferes with the lymphocyte transformation only when the cells are already activated with PHA.     

Effect of a streptococcal allergen on the rosette-forming capacity of the T-lymphocytes in erysipelas patients

The reaction of antigen-dependent E-rosette formation with the hemolytic streptococcal antigen in erysipelas patients is indicative of the ambiguous role of the specific immunological transformation of the body in respect of the infective agent antigens in different clinical forms of the disease and is of prognostic importance as regards the chronic transformation of the infectious process and the development of the relapses of the disease.     

Comparative study of the virulence of the Shigella isolated from bacterial carriers and from acute dysentery patients]

The authors present the results of study of the virulence of shigellae isolated from carriers and patients suffering from acute dysentery, on a continuous culture of E1 and Hep-2 cells. The virulence of shigellae isolated from carriers displayed no significant difference from the virulence of shigellae isolated from patients with a mild and moderately severe forms of dysentery. In the patient's organism shigellae were capable of retaining the initial virulence for a long time, despite the treatment and the influence of the macroorganism's protective factors. The authors believe that this was connected with the capacity of Shigellae to parasitic life in the cells of human intestinal epithelium, and, apparently, played a definite role in the formation of carrier state.     

Mechanism of virus-induced immunopathology. I. The reactivity of immunocompetent cells in acute herpetic infection in an experiment

The state of general and specific responsiveness of thymocytes and splenocytes in non-inbred white mice has been studied in the reaction of lymphocyte blast transformation under the influence of phytohemagglutinin (PHA) and herpes simplex virus. Experimental herpetic encephalitis has been shown to give rise to the development of pronounced immunosuppression, which is confirmed by a considerable decrease (P less than 0.05) in the levels of the PHA- and virus-induced transformation of thymocytes and splenocytes in infected mice in comparison with the similar transformation characteristics of intact lymphocytes. The inhibition of the general and specific responsiveness of T-lymphocytes has been found to be one of the possible mechanisms of immunosuppression in acute herpes infection.     

Effectiveness of the antibacterial therapy of patients with various forms of dysentery]

The analysis of the efficacy of various methods of antibacterial therapy in 1382 patients with obliterated, light and moderately severe forms of dysentery as compared to the patients subjected only to patholgenic therapy showed that broad spectrum antibiotics lost their role in treatment of such patients. Furazolidon proved to be one of the most active drugs. The authors suppose that treatment of patients with obliterated forms of dysentery with antibacterial drugs is not obligatory and is indicated only in cases with repeated isolation of the causative agent.     

Increasing the effectiveness of antibiotic therapy by correcting immunologic disorders with vitamin A in patients with brucellosis

Experimental studies and clinical trials were performed on possible increase of antibiotic therapy efficacy in brucellosis patients by correction of the immunity disorders with vitamin A. It was experimentally shown that vitamin A increased cellular immunity and accelerated sanation of guinea pigs sensitized with Brucella abortus 19 BA. The clinical trials demonstrated that the use of vitamin A in a dose of 33,000 IU thrice a day for 10 to 12 days during the complex treatment of patients with acute (36 persons) and subacute (57 persons) brucellosis lowered the average period of manifestation of the disease clinical signs and formation of the antibodies, increased the skin allergic sensitivity, the lymphocyte blast cell transformation, the total number and subpopulations of the active T-cells, theophylline-resistant lymphocytes, phagocytic and metabolic activity of neutrophils, showed 1.5- and 2-fold increased in the frequency of the infection transformation into a chronic process in patients with acute or subacute brucellosis, respectively.     

The immunological diagnosis of peptostreptococcal infection

The work deals with different methods for the diagnosis of anaerobic streptococcal infection, experimentally tested and clinically approved in the examination of children with acute pneumonia. The passive hemagglutination test, the immune rosette formation test and the specific lymphocyte blastogenesis test were used for diagnosis. As sensitin and antigen, the preparation of the peptostreptococcal bacterial mass was used. The diagnostic titer of antibodies to anaerobic streptococci was 1:160. A twofold and greater increase in the number of antigen-dependent rosette-forming lymphocytes and in specific blast transformation in the presence of the diagnostic antibody titer confirms the etiological role of anaerobic peptostreptococci in the development of pneumonia in children.     

In vitro induction of antigen specific antibody synthesis and proliferation of T lymphocytes with acellular pertussis vaccines, pertussis toxin and filamentous haemagglutinin in humans

The in vitro response of human B- and T-lymphocytes to the acellular vaccines JNIH-6 (containing pertussis toxoid and filamentous hemagglutinin), and JNIH-7 (containing pertussis toxoid), and to the purified components JNIH-4 (filamentous hemagglutinin) and JNIH-5 (pertussis toxin) was investigated. Pertussis toxoid and filamentous hemagglutinin induced specific Ig synthesis in vitro in lymphocytes obtained from convalescent pertussis patients as target cells. The antigen-dependent Ig production was demonstrated in lymphocyte culture supernatants by ELISA techniques and by a chinese hamster ovary cell toxin neutralization assay. Particularly with JNIH-4, -6 and -7, high antibody titers were obtained. At optimal antigen concentrations a marked lymphocyte blast transformation was found in lymphocyte cultures from whooping cough patients, but not in cultures of lymphocytes obtained from healthy volunteers. At high concentrations native pertussis toxin as well as the B oligomer (S2-5) of the toxin induced a strong proliferation of patient as well as control lymphocytes, indicating non-specific mitogenic activity. At lower concentrations lymphocyte blast transformation was seen in patient cultures only, which indicates an antigen-specific T-cell response. The A protomer (S1), dimer 1 (S2 + 4) and dimer 2 (S3 + 4) induced proliferation of patient lymphocytes, which demonstrates the presence of T-cell epitopes on these peptides. The in vitro B-cell response and the lymphocyte blast transformation assay are both useful tools for estimating the potency of acellular pertussis vaccines in man. Spontaneously acquired and vaccine induced immunity to Bordetella pertussis can be investigated at the level of B- and T-lymphocytes.     

Antigenic heterogeneity in patients with reactions in borderline leprosy.

Fifteen patients with borderline leprosy who developed "reversal" reactions were studied from the inception of treatment. Thirteen showed an appreciable increase in lymphocyte transformation (LT) when preparations of Mycobacterium leprae were used as antigen. The LT responses to either "whole" or "sonicated" preparations of the bacillus in these 15 patients and in nine others also in reaction correlated with the clinical presentation. Those with skin disease predominating in the reaction showed an appreciable increase in LT when whole M leprae was used as antigen. Those with nerve disease predominating showed an increase with sonicated M leprae. In those with both skin and nerve disease there was an increase with both antigen preparations. The ratios of the LT test results (whole to sonicated M leprae) showed highly significant differences between the three groups.     

In vitro study of macrophages from patients with sarcoidosis

Cultures of leukocytes from patients with sarcoidosis were stimulated with Kveim antigen or PPD-S and the macrophage reactions were compared with those obtained under similar conditions with leukocyte cultures from healthy tuberculin reactors. The non-stimulated control cultures of Kveim-positive sarcoid patients showed signs of macrophage activation, namely an increase in cytoplasmic pyroninophilia in small and enlarged macrophages, together with a higher than normal rate of lymphoblast transformation. In the presence of Kveim antigen this activation was more marked, even in cases where the blast response to the antigen was of low magnitude. Stimulation with tuberculoproteins produced a slight activation of macrophages only in cultures from Kveim reactors who had also shown positive tuberculin reactions.These observations suggest that the study of lymphocyte transformation together with that of macrophage activation will improve the immunocytological assessment of sarcoidosis.     

A comparison of the dynamics of changes in large intestinal mucosa of acute dysentery patients with detection of a specific antigen in urine

The relationship between the elimination of shigellae from the body with urine and the dynamics of morphological changes in the mucous membrane of the large intestine of acute dysentery patients at the early convalescence period has been studied. The persistence of Shigella antigen in acute dysentery patients and its elimination with urine is, as a rule, accompanied by local immune cell reaction in the mucous membrane of the large intestine. The antigen circulating in the body is a factor contributing to the inflammatory process in the intestine, the amount of the eliminated antigen being higher in pronounced inflammations of the intestinal mucosa than in residual inflammatory phenomena. A group of patients (3 persons) releasing Shigella O-antigen with urine and having the inflammatory process in the large intestine, but showing no signs of local immune reaction in the lymphoid tissue of the large intestine, has been found. The reactivity of the lymphoid tissue of the intestine in such patients is a risk factor contributing to the development of prolonged dysentery or chronic postdysenteric colitis.     

State of cellular immunity and phagocytes in guinea pigs infected with Mycoplasma pneumoniae

The work presents the results of the study of experimental Mycoplasma pneumoniae infection in guinea-pigs. A considerable increase in the engulfment of mycoplasmas and blood leukocytes was found to occur on days 14-28 after the infection. The correlation between the degree of the engulfment of mycoplasmas by macrophages and the activity of lymphocytes in the reaction of blast-cell transformation with phytohemagglutinin and Mycoplasma antigen was observed. The peculiarities of the course of infection in sensitized animals were revealed. Virulent Mycoplasma pneumoniae strains were found to have a toxic effect on the lymphocytes and macrophages of guinea-pigs. This effect was neutralized by specific antiserum. The importance of these facts for the pathogenesis of Mycoplasma infection is discussed.     

Monoclonal antibodies against the alpha-chain of human lymphocyte function-associated antigen-1

The goal of the investigation was to further characterize ICO-II monoclonal antibodies. ICO-II have been shown to block NK activity of mononuclear cells from the blood of healthy donors against K-562 and Molt-4 target cells, cytotoxic T lymphocytes induced in a 7-day mixed lymphocyte culture, the reaction of lymphocyte blast cell transformation to phytohemagglutinin and the formation of En-rosettes. The molecular weight of the antigen detected by ICO-II is 180 KD. ICO-II are shown to detect alpha-subunit of human lymphocyte function-associated antigen-I (LFA-I).