The historical experience and practice of fight against tuberculosis in country which is one of the high drug resistant-tuberculosis (DR-TB) burden countries in European Union (EU).

Despite considerable efforts and quite early initiated anti-tuberculosis (TB) actions, Lithuania still remains one of the European Union (EU) countries with the highest tuberculosis rates, especially multidrug-resistant (MDR) TB. According to the European Centre for Disease Prevention and Control, in 2016, 58 994 cases of TB were reported in 30 EU/European Economic Area (EEA) countries. MDR TB was reported for 3.7% of 36 071 cases with drug susceptibility testing results and continues to be highest in the three Baltic countries - Estonia, Latvia and Lithuania. In this article we present the Lithuanian anti-TB action history review and comparison with other countries in this area of action. Literature review was performed by using documents available in the Martynas Mazvydas Library's resource, articles of foreign authors and archival materials. According to archaeological studies, tuberculosis was common in Europe including Lithuania in the Middle Ages. Tuberculosis reporting started in Lithuania in 1926. The first tuberculosis sanatorium in Lithuania was opened in 1891. Patients were treated with sun bathing procedures, fresh air and sunlight. Later the treatment included pneumothorax, toracocaustic, toracoplastic, treatment with gold products and other procedures. Lithuania introduced directly observed treatment, short course therapy (DOTS) in 1999, and since 2007 it has been working in accordance with the requirements of this strategy.

Evaluation of different laboratory methods for diagnosis of pig chlamydiosis in Lithuania.

Pig chlamydiosis is antrophozoonosis caused by Chlamydophila abortus. Chlamydias (C type) are widely found in nature and can infect humans, domestic and wild mammals, and 139 types of birds. The peculiar feature of chlamydias is the tropism to different tissues, organs and organisms. In 2502 pig blood sera tests from Lithuanian farms, anti-chlamydia complement binding (CB) antibodies were detected in 192 cases (7.7%). Serological tests showed the following (C type) chlamydia bearing regions: 22.0% Mazeikiai district, 17.2%--Kaisiadorys district, 13.5%--Panevezys district, 12.3%--Vilkaviskis district. Rare incidence of the disease was found in Siauliai district 1,2% and Klaipeda district 2.5% farms. The highest antibody titers in blood serum tests were found in Joint Stock Company (JSC) "Krekenava" and "Vejine", i.e. 1:128 and 1:64, respectively. The following methods for the study of pig chlamydiosis were used and comparatively evaluated: complement binding reaction (CBR), direct immunofluorescence (DIF), imunoenzyme assay (IEA), indirect immunofluorescense (IIF), micro immunofluorescense (MIF), polymerase chain reaction (PCR) and cell culture (CC) test. PCR method was found to be more sensitive and reliable compared to imunoenzyme assay, but the latter is more economic especially for screaning. In pigs with the clinically expressed symptoms, 108 pigs infected with chlamydia were detected. CB assay revealed the infection rate from 3.4% to 7.9% in piglets, sows and boars. The highest level of chlamydia infection was detected in fatteners (17.6%). Seroepizootic study of pig chlamydiosis revealed the different infection rate in the animals investigated. The highest chlamydia infection risk is in winter (10.4%) and the lowest--in summer (2.8%).

Chronic hepatitis C: T-helper1/T-helper2 imbalance could cause virus persistence in peripheral blood.

Interferon gamma (IFN gamma) and interleukin 4, 10 and 12 (IL-4, -10, -12) production was measured in whole peripheral blood (WPB) and peripheral blood mononuclear cells (PBMC) of 10 chronic hepatitis C (CHC) patients. The level of IFN gamma in supernatants in mitogen-activated WPB was lower than in healthy donors. IL-10 served as a possible downregulative factor for IFN gamma, since its spontaneous IL-10 production was enhanced in CHC. Neutralization of IL-10 partly restored IFN gamma response in CHC patients. Recombinant IL-12 (rIL-12) also enhanced IFN gamma of CHC patients, but IL-12 production was decreased in CHC. Thus, IFN gamma production deficiency in CHC patients is secondary to blockage by high levels of IL-10-impaired IL-12 production.

A rapid ELISA test for detection of human paraproteins.

A rapid ELISA test for detection, characterization and quantification of human paraproteins was developed. The proposed method is a sandwich ELISA, where the capture antibody is specific for a given heavy chain (gamma, alpha or mu) and the labelled antibody is specific either for kappa or for lambda light chain. Both standard and patient sera are tested with all six possible antibody combinations. Each paraprotein produces a significant increase in titre (as compared with standard) only when tested with the relevant pair of antibodies. This enables the determination of the isotype and light chain type of the paraprotein and the evaluation of its relative quantity in patient serum. The accuracy of the assay (relative deviation) varies from 0.04 for gamma lambda to 0.19 for alpha kappa. The cut-off values for each type of polyclonal immunoglobulin were determined with 200 healthy donor sera. 103 patient sera were analysed. ELISA data are in good agreement with M-component and other clinical data.

Immunochemical study of human immunoglobulin G Fc region.

Fifteen murine monoclonal antibodies (mAb) were produced that reacted with conformational epitopes present on the Fc portion of all human IgG subclasses (PAN-Fc). Inhibition and sandwich ELISA were used to elucidate overlapping and distinct epitopes. The epitopes were aligned to form a continuous "chain" of overlapping determinants from the CH2-CH3 interface to the direction of hinge. The five more hinge-proximal epitopes demonstrated high lability both in competitive and sandwich assays, being completely or partially destroyed when any of these 15 other mAb bound the IgG first. Heat-inactivation of IgG caused full disruption of these epitopes. In contrast, epitopes situated at the opposite distal of the "chain" were more stable and mAb binding could only be affected by occupying an overlapping epitope. Under heat-inactivation these epitopes were affected, but not completely destroyed. Human IgG class anti-DNA autoantibodies were bound to insolubilized dsDNA and their reaction with PAN-Fc mAb was studied. mAb titration plots on IgG and dsDNA-IgG were compared. Five epitopes proved to be altered by antigen (dsDNA) binding. Two of these were the labile hinge-proximal epitopes and the other three were situated near the CH2-CH3 interface. Cross-reactivity of mAb with xenogeneic IgG was also studied. An "epitope map" of the crystallographic model of human IgG Fc portion drawn was based on these experimental data and printed matter, concerning the location of subclass-specific amino acids and homology regions of human and animal IgG.

Partial characterization of bovine leukemic cell populations.

Peripheral blood lymphocytes (LL) from cows with chronic lymphocytic leukemia (CLL) have been studied using a number of surface markers. Cell populations were obtained by partial enrichment and depletion using Sephadex G-200-anti-F(ab')2 and Sephadex G-200-anti-LL-Ig immunoadsorbent columns. It was shown that cell populations having different markers, i.e., surface immunoglobulins (sIg) and leukemia-associated antigens (LAA), are characterized by similar parameters. Thus the adherent cells obtained by both fractionation methods formed 1.1-4.0% of rosettes with sheep red blood cells, while 90.6-94.3% were sIg positive cells. The cytotoxicity test (CTT) performed with anti-B and anti-LL sera indicated that a vast majority of adherent cells reacted to the sera. Thus the adherent cells represent a population with a high percentage of B-cells.

Establishment of a cell line from leucocytes of a cow with chronic lymphocytic leukemia.

A cell line was established from blood leucocytes of a cow with chronic lymphocytic leukemia. The leucocytes were cultured with conditioned medium (culture fluid of mouse cell line L). In vitro cell transformation was demonstrated by adaptation to permanent growth, modification of cell morphology, the alteration of cell surface phenotype, kinetic behaviour and the loss of the euploid stability of the cell karyotype. Ultrastructural studies showed rather a uniform cell pattern in a culture population heterogeneous for degree of cell vacuolization. A wide variation in the expression of surface markers in cells was demonstrated by E-, EA- and EAC-rosetting. In suspension culture the cell population was found to be sIg negative. Expression of leukemia-associated antigens by a fraction of the cultured cells was evidenced by a cytotoxic technique using complement and heterologous antisera against bovine leukemic lymphocytes, absorbed with normal lymphoid cells. Virus-like particles and BLV antigens were not identified. Culture cells failed to show spontaneous or antibody-dependent killer cytotoxicity. Comparison with blood lymphocytes of healthy and leukemic cattle was done. The established culture should be useful as a model for experimental immunology and oncology.