Lymphocyte stimulation test in the diagnostics of adverse reactions to antituberculouse line 1 and 2 drugs.

Adverse drug reactions to anti-TB drugs (ADR) are found in 6-20% of patients and have various clinical manifestations and are detected in the lymphocyte stimulation test (LST), recorded by the incorporation on H3 thymidine, but nowadays it has significant limitations. We used LST with WST-1 reagent to detect ADR to the main 1-st and 2-nd line antituberculosis drugs in 11 tuberculosis patients who had ADR (6 - hepatotoxic reaction, 3 - blood eosinophilia and 2 - with joint pain syndrome). 6 people with tuberculosis contacts made up the control group. LST evaluation with WST-1 showed that in patients with a hepatotoxic reaction, the SI index was>2 and exceeded the values in the control group (3.28±0.59, 95% CI-1.16 and 0, 74±0.16, 95% CI - 0.31, respectively) upon stimulation of cell cultures with rifampicin alone but not with other drugs. Cell cultures stimulated with the PHA mitogen have SI >2 in ADR patients (mainly with hepatotoxic reactions). Control group SI was <2 (4,93±0.53, 95% CI - 1, 04 and 1.97±0.3, 95% CI - 0.59, respectively). We have not detected PPD-L cell cultures stimulation with WST-1 reagent both in the group of patients with ADR and the control group. In patients with eosinophilia and joint pain syndrome SI was low for all studied drugs and did not differ from the control group. The sensitivity of the LST test with WST-1 reagent is not sufficient to determine ADR to anti-TB drugs.

[Tuberculous IgE antibodies. Part II. Study of its concentrations in different forms of tuberculosis]. / Protivotuberkuleznye IgE-antitela. Chast' II. Issledovanie kontsentratsii pri razlichnykh formakh tuberkuleza.

Tuberculosis-afflicted lung are infiltrated by two functionally types of lymphocytes, which presumably counteract with each other by producing proinflammatory (type 1) and anti-inflammatory (type 2) cytokines. It is held that irregular sequestration of antigen into different compartments of the lung may lead to preferential activation of T-helper 1 or T-helper 2 lymphocytes. Unlike IgE antibodies, specific tuberculosis IgE antibodies are seen only in tuberculosis infection. The mean values of IgE antibodies in tuberculosis (7.661 +/- 0.849 IU/ml) are significantly greater than those in other pulmonary diseases (1.768 +/- 0.116 IU/ml). Low concentrations of tuberculosis IgE antibodies in persons with a marked hyperergic response to tuberculin (1.808 +/- 0.097 IU/ml) are of importance. Significant concentrations of mycobacterial IgE antibodies are mainly detected in fibrocavernous (14.56 +/- 1.11 IU/ml), infiltrative (10.10 +/- 1.08 IU/ml), peripheral lymph nodal (10.53 +/- 1.09 IU/ml) tuberculosis rather than intrathoracic lymph nodal tuberculosis (4.555 +/- 0.340 IU/ml). There is a particularly considerable increase in specific IgE antibodies in a phase of decay (15.98 +/- 1.64 IU/ml) and infiltration (12.66 +/- 1.08 IU/ml). These groups also show a concurrent rise in tuberculosis IgG antibodies, which nevertheless disagree with the increase of IgE (the correlation coefficient is 0.599).

[Anti-tuberculous IgE antibodies. I. Immunodominant antigens]. / Protivotuberkuleznye IgE-antitela. I chast'. Immunodominantnye antigeny.

It is widely accepted that protection against tuberculosis is provided by the formation of type 1 immune response, which is characterized by the production of IFN-gamma and IL-2. However, type 2 antimycobacterial immune response is also present: specific IgE antibodies that are IL-4 dependent, are usually found in tuberculosis patients. There is elevated production of type 2 cytokines in some cases. Thus, both types of an immune response can simultaneously develop, probably counteracting with each other. It is unknown which of mycobacterial antigens are capable of inducing a preferential type 2 response. To detect these antigens, the authors studied tuberculosis IgE antibodies in the sera of 500 tuberculosis patients by using the ELISA assay with ultrasonic disintegrated M. Tuberculosis H37Rv (sonicate). Antigens recognized by IgE antibodies were found to be localized in the cell wall of mycobacteria. The IgE-response was specific since the sera did not react with the antigens of atypical mycobacteria and other bacterial species.

[Experience with kipferon use in the treatment of patients with pulmonary tuberculosis]. / Opyt primeneniia kipferona pri lechenii bol'nykh tuberkulezom legkikh.

Kipferon that is a combination of recombinant human (2-interferon and a complex of immunoglobulins G, M and A, was used in suppositories as an auxiliary agent in the routine chemotherapy in 36 new cases of pulmonary tuberculosis. A control group included 19 patients identical in sex, age, and the pattern of pulmonary tuberculosis. The clinical, X-ray, and laboratory indices (primarily cellular immunity) were studies before and 1 and 3 months after treatment. The beneficial effect of kipferon was manifested by a more rapid arrest of symptoms of total intoxication eliminated after 2 weeks in 39% of patients in the experimental group and only in 21% in the controls. Normalization of blood parameters occurred following a month in 58.3 and 47% of patients, respectively. Mycobacteria tuberculosis disappeared in the sputum smears following a month of treatment in 62% of those isolating bacteria in the experimental group and only in 37.5% in the controls (P > 0.1; t = 1.6). Positive lung X-ray changes as resolved infiltration, the reduction and closure of caverns were more pronounced in the patients of the experimental group. The most characteristic change in the parameters of cellular immunity during kipferon was a short (as long as 1-1.5 months) decrease in RBT to FGA, which was noted in 47% and 6.7% of patients in the experimental and control groups, respectively (P < 0.01) and which was followed by an increase in the count of CD8+ cytotoxic lymphocytes. This is indicative of the enhancement of these mechanisms of immunity and a reduced need of enhancing or maintaining the activity of proliferative reactions of immunocompetent cells under the conditions of a favourable influence on the course of tuberculous infection.

[Immunology of tuberculosis: current status]. / Immunologiia tuberkuleza: sovremennoe sostoianie problemy.

The paper outlines the high-priority trends in basic and applied investigations of antituberculous immunity. It presents the results obtained from studies of the genetic mechanisms responsible for the control of susceptibility in experimental tuberculosis, from analyses of novel chemical and recombinant tuberculosis vaccines, from examinations of the role of immunological memory in the disease. While describing the results of tuberculosis immunodiagnosis, particular emphasis is laid on its major aspects: on the differential diagnosis of tuberculosis and other lung diseases and on the screening of high-risk group populations for disease progression in order to made further in-depth studies. Current aspects of evaluating the immune status and nonspecific responsiveness in patients with tuberculosis are considered.

[The activity of immunocompetent cells and macrophages in pulmonary sarcoidosis (in comparison to tuberculosis)]. / Aktivnost' immunokompetentnykh kletok i makrofagov pri sarkoidoze legkikh (v sravnenii s tuberkulezom).

Composition of immunocompetent cells and production of cytokins (IL-1, IL-2, IL-6 and PNO) by stimulated and nonstimulated cells of blood and bronchoalveolar lavage (BAL) were evaluated in 35 and 25 sarcoidosis and tuberculosis patients, respectively. Comparable differences were recorded in concentration of T-cells and T-helpers versus BAL in sarcoidosis and tuberculosis as well as in intensity of cytokin production by mononuclears obtained from various sources.

[The immunomodulating action of cyclosporin A in vitro and in vivo on the lymphocytes of patients with alveolitis]. / Immunomoduliruiushchee deistvie tsiklosporina A in vitro i in vivo na limfotsity bol'nykh al'veolitami.

The paper presents the first results of using cyclosporin A (CsA) to treat lymphocytes during their extracorporeal immunomodulation (EIL) in patient with fibrotic alveolitis of various etiology. Two-hour lymphocytic incubation in the medium containing 0.1-10 micrograms per ml of CsA was sufficient for CsA to show its in vitro immunosuppressive effect, which resulted in a substantial inhibition of a proliferative lymphocytic response to mitogens and antigens. Administration of CsA-treated lymphocytes induced no profound structural changes in lymphocytic subpopulations (CD3, CD4, CD8), but it was followed by a reduction in the baseline high proliferative lymphocytic response to PHA. The clinical effect, alveolitis alleviation was noted in all patients. It is suggested that clinical effects may be produced by a local concentration of the treated lymphocytes and their transferred CsA as well.

[Use of bronchovaxom in the treatment of patients with chronic obstructive bronchitis and bronchial asthma]. / Primenenie bronkhovaksoma v lechenii bol'nykh j khronicheskim obstruktivnym bronkhitom i bronkhial'noi astmoi.

Polyvalent vaccine bronchovaxom (BV) was studied for efficacy against chronic obstructive bronchitis and bronchial asthma exacerbation. Good results were reported: reduction of disability duration, number of recurrences, cough intensity, discharged sputum. Laboratory tests discovered that BV brought about IgE decrease, IgA, T3, T4/T8 increase in bronchoalveolar lavage.

[Cytokines in tuberculosis]. / Tsitokiny pri tuberkuleze.

The paper presents the data available in the literature and the authors' own findings concerning the production of cytokines, such as interleukins 1, 2, 4, 6, and 8, interferons and tumor necrosis factor, in patients with different stages of tuberculosis. A relationship between the production rate of some cytokines and the stage of the disease, the extent of the process, chemotherapeutical efficiency and other clinically important factors is discussed. The prospects of further investigations in this area are dealt with.

[Cytokines in immunopathogenesis of pulmonary tuberculosis]. / Tsitokiny v immunopatogeneze tuberkuleza legkikh.

Production of IL-1, IL-2, gamma-interferon and PNO was measured in 46 patients with various forms of pulmonary tuberculosis. Cytokins quantitation was conducted biologically, using radioimmunoassay and enzyme immunoassay in supernatants of cellular structures (blood lymphocytes). There were high levels of IL-1 and PNO, reduced production of IL-2 and gamma-IFN by stimulated blood mononuclears. In effective chemotherapy (3-4 months) production of IL-1 tented to a decline, while that of IL-2 and gamma-IFN went up.

[Phenotypic and functional analysis of blood lymphocytes and pathological focus in patients with pulmonary tuberculosis]. / Fenotipicheskii i funktsional'nyi analiz limfotsitov krovi i patologicheskogo ochaga u bol'nykh tuberkulezom legkikh.

A subpopulation study of lymphocytes concentrating in cavernous, lymph node tissues and blood from patients with fibrous-cavernous tuberculosis of the lungs showed that in spite of high CD3+ and CD4+ cell levels in the cavern, the response of these cells to mitogens may be weaker than blood lymphocyte response. IL-2 production by cavern lymphocytes varied and was unrelated to proliferative response of these cells to M. tuberculosis antigens.

[Decreased count of small lymphocytes in the blood of patients with malignant bone tumors]. / Snizhenie soderzhaniia malykh limfotsitov v krovi bol'nykh so zlokachestvennymi kostnymi opukholiami.

Blood small lymphocyte (not exceeding 7.5 micron in diameter) counts obtained from patients with malignant bone tumors in the course of primary examination were 40-75% those in healthy subjects. The said changes were registered only in some patients with osteoblastoclastoma; they were not observed in cases of trauma, osteomyelitis, sepsis, spontaneous osteolysis and chronic synovitis. The study failed to establish a correlation between blood small lymphocyte count, on the one hand, and concentrations of total, stable and active T-lymphocytes as well as autologous E-rosettes, on the other. In the course of separation of lymphocytes in percoll density gradients, small lymphocytes concentrated in high density fractions. Purified small lymphocytes of healthy subjects appeared to be mainly T-lymphocytes, particularly, "activated" ones. Proliferative response to PHA and production of interleukin-2 in cell cultures showing high levels of small lymphocytes were higher than in those with moderate or low concentration of the said cells. Small lymphocytes are considered to be a special subset of T-cells which exhibit high functional activity and may be identified only morphologically. Lowered counts of these cells are attributed to neoplastic growth.

[Comparative analysis of immune response indices in lung cancer]. / Sravnitel'nyi analiz pokazatelei immunologicheskoi reaktivnosti pri rake legkogo.

In 43 patients with cancer of the lung, a number of immunologic indexes were evaluated including lymphocyte response to PHA, lymphokine production, serum immunoglobulin levels and the regulatory effect of sera and T-lymphocytes on lymphokine production. No significant deficiency in T- and B-lymphocytes was registered in patients with stage I-III cancer, nor any rise in TG cell level was in evidence. The response to PHA was low; however, the capacity to produce lymphokines in the presence of allogeneic or autologous tumor antigens was unimpaired. Purified T- and TG-cells isolated from blood or tumor tissue suppressed immune response in leukocyte migration inhibition test. However, this effect was not related to the increase in levels of these cells in the circulation. The autologous sera of practically all tumor-sensitized patients exerted a blocking effect in vitro. This effect was related neither to levels of serum immunoglobulins, nor to those of blood TG cells.

[Comparative analysis of immunoreactive indices in malignant and benign bone tumors]. / Sravnitel'nyi analiz pokazatelei immunologicheskoi reaktivnosti pri zlokachestvennykh i dobrokachestvennykh opukholiakh kostei.

The paper discusses the results of a study of immunologic surveillance in patients with bone tumors. T-lymphocyte deficiency was found in 21-32% of cases only, while all patients showed changes in the level of some T-cell subpopulations and their ratio. Patients with malignant tumors (mostly osteosarcoma) revealed a lowered response to PHA and a high frequency of sensitization to tumor antigens. In those cases, serum, blood--circulating T-cells and tumor--infiltrating lymphocytes showed a suppressor activity. A moderately decreased response to PHA, a low frequency of sensitization to tumor antigens and a stimulating activity in serum, blood--circulating T-cells and lymphocytes were registered in cases of benign tumors (mainly giant cell tumors). Suppressor activity of blood T-lymphocytes was matched by a high level of TG cells, while stimulating effect--by a low one.

[Lymphocyte subpopulations in malignant bone tumors and their relation to the immune response to the tumor]. / Subpopuliatsii limfotsitov v kostnykh slokachestvennykh opukholiakh i ikh sviaz' s immunnym otvetom na opukhol'.

Comparative studies of the subpopulations of lymphocytes, isolated from the tumor tissue and blood of 15 osteosarcoma patients, showed that the total lymphocyte count in the neoplasms ranged from 0 to 19% of the total cell population. In various patients tumor lymphocytes were represented either mainly by T cells (up to 80% of lymphocytes) or by "null" cells lacking any surface markers. The population of lymphocytes with surface membrane immunoglobulins with complement and Fc receptors was usually low although the tumors contained up to 90% of nonlymphoid cells with the same markers. Lower (less than 60%) counts of blood T lymphocytes were found only in one third of osteosarcoma patients. The immune response of blood lymphocytes against living autologous tumor cells was found in 85% of the patients, and its intensity depended on the content of T lymphocytes in tumor but not in blood.

[Rheumatoid factor in oncological patients]. / Revmatoidnyi faktor u onkologicheskikh bol'nykh.

In sera of 20% of patients with tumors of different localization and histological type the rheumatoid factor (RF) was found, which more frequently occurs in patients with soft tissue tumors as compared with bone tumors. The blocking properties of the sera revealed by RBT to PHA and by RIML would show no correlation with the occurrence of RF in them. Water-salt extracts of some tumors contain RF, which occurrence correlates well with the concentration of tumor cells having receptors for Fc-site of IgG on their surface.

[In vitro immune response of patient lymphocytes to live autologous tumor cells]. / Immunnyi otvet in vitro limfotsitov bol'nykh na zhivye kletki autologichnoi opukholi.

In 87% of patients with malignant tumors and in 24% of patients with benign tumors there was found the in vitro immune response of lymphocytes to autologous tumor cells. Most of the autologous sera influenced the immune lymphocytes: those of malignant tumor patients usually blocked while those of benign tumor patients stimulated the immune reaction. Both the blocking and stimulating serum activities could be removed by absorption of the sera with living autologous tumor cells. Following the absorption intact sera of some patients with malignant tumors obtained the blocking capacity.

[Nature of the blocking action of the sera of oncological patients on T-lymphocytes]. / Izuchenie prirody blokiruiushchego deistviia syvorotok onkologicheskikh bol'nykh na T-limfotsity.

Some sera of tumor patients render a blocking effect on the activity of autologous and allogenic T lymphocytes in blasttransformation test to PHA and spontaneous rosette-formation. The activity that block spontaneous rosette-forming reaction is directed against the population of postthymic peripheral lymphocytes but not that of thymocytes. No correlation was found between the serum activity, which block RSP and RBT, and antibodies to PHA, lymphocytotoxins, C-reactive protein and the rheumatoid factor.