Induction of arthritis in BALB/c mice by cartilage link protein: involvement of distinct regions recognized by T and B lymphocytes.

Both type II collagen and the proteoglycan aggrecan are capable of inducing an erosive inflammatory polyarthritis in mice. In this study we provide the first demonstration that link protein (LP), purified from bovine cartilage, can produce a persistent, erosive, inflammatory polyarthritis when injected repeatedly intraperitoneally into BALB/c mice. We discovered a single T-cell epitope, located within residues 266 to 290 of bovine LP (NDGAQIAKVGQIFAAWKLLGYDRCD), which is recognized by bovine LP-specific T lymphocytes. We also identified three immunogenic regions in bovine LP that contain epitopes recognized by antibodies in hyperimmunized sera. One of these B-cell regions is found in the most species-variable domain of LP (residues 1 to 36), whereas the other epitopes are located in the most conserved regions (residues 186 to 230 and 286 to 310). The latter two regions contain an AGWLSDGSVQYP motif shared by the G1 globulin domain of aggrecan core protein, versican, neurocan, glial hyaluronan-binding protein, and the hyaluronan receptor CD44. Our data reveal that the induction of arthritis is associated with antibody reactivities to B-cell epitopes located at residues 1 to 19. Together, these observations show that another cartilage protein, LP, like type II collagen and the proteoglycan aggrecan, is capable of inducing an erosive inflammatory arthritis in mice and that the immunity to LP involves recognition of both T- and B-cell epitopes. This immunity may be of importance in the pathogenesis of inflammatory joint diseases, such as juvenile rheumatoid arthritis, in which cellular immunity to LP has been demonstrated.

Arthritis induced by proteoglycan aggrecan G1 domain in BALB/c mice. Evidence for t cell involvement and the immunosuppressive influence of keratan sulfate on recognition of t and b cell epitopes.

Our previous work showed that the proteoglycan aggrecan can induce erosive polyarthritis and spondylitis in BALB/c mice, and that the G1 domain of the proteoglycan aggrecan (G1) is the arthritogenic region. In this study, two T cell epitopes residing on G1 within residues 70-84 (peptide G5) and 150-169 (peptide G9) were identified using synthetic peptides and aggrecan-specific T cell lines. Two G1-specific T cell hybridomas exclusively responded to peptide G5. When the G5-specific T cell line was injected intraperitoneally into BALB/c mice, it induced acute inflammatory arthritis in joints, but only in those that had been injected with the epitope recognized by these T cells. Furthermore, we also demonstrate that the keratan sulfate chain(s) (KS) on G1 possess immunosuppressive properties with respect to T and B cell epitope recognition. T cell lines that recognize both G1 and peptide G5 show an increased response to G1 after KS is removed. Antibodies in hyperimmune sera of mice immunized with G1 show increased epitope recognition (quantitative and qualitative) after KS removal before immunization. These studies reveal that a T cell line specific to an epitope on the G1 domain of aggrecan, also recognizing a corresponding mouse G1 epitope, can induce arthritis by adoptive transfer and homing to the intraarticular epitope, thereby implicating T cells in arthritis development caused by immunity to the G1 domain of aggrecan. Moreover, the presence of KS on G1 can inhibit arthritis development by suppressing T and B cell epitope recognition.

[Modern immunologic tests in the diagnosis of Lyme borreliosis]. / Savremeni imunoloski testovi u dijagnostici lajm borelioze.

In this work we described the results that were obtained using various immunodiagnostic assays for the detection of lyme borreliosis. Sera of the patients that were in acute or chronical phase of the disease were analysed in indirect immunofluorescent, immunoenzyme and immunoblot assays which were prepared and carried out in our laboratory. As a control for the validity of these investigations, we used sera of the healthy people, as well as of the patients suffering of lues or rheumatoid illnesses. Results that we obtained pointed out the factors responsible for the nonspecific reactions in indirect immunofluorescent and immunoenzyme test. The advantage of the immunoblot analysis in detecting lyme borreliosis is described in this work.

[Conditions required for chemical binding of Dactylis glomerata grass pollen to inert stable carriers]. / Ispitivanje uslova za hemisjsko vezivanje polena trave dactylis glomerata na inertne stabilne nosace.

The results of the direct RAST (Radio-Allergo-Sorbent-Test) procedure used to standardise own-prepared pollen extract of the Dactylis glomerata grass are presented. Based on the performed standardisation, a system was set up for binding allergens to the stable inert carriers. The results of examination of the quality of D. glomerata allergen, bound covalently to paper disc, are presented also. Evaluation was made in INEP laboratories by comparing the D. glomerata allergen-bound discs and the corresponding discs of Phadebas, Pharmacia Uppsala, confirming a high degree of correlation in the direct RAST procedure.

[Use of the ELISA immunoenzyme test and indirect immunofluorescence in the diagnosis of a toxoplasmosis epidemic]. / Primena imunoenzimskog testa ELISA i indirektne imunoflurescencije u dijagnozi epidemije toksoplazmoze.

Etiologic agent of lymphadenitis outbreak in one our military environment was detected by two serologic tests: ELISA (enzyme-linked immunosorbent assay) and indirect immunofluorescence test (IIF) both specific for IgM and IgG antibody detection in toxoplasmosis diagnosis. IgM-ELISA analysis of the first serum sample taken from a group of 79 soldiers with lymphadenitis was positive in 65 examinees (82.28%) and IgG-IIF technique in 52 (65.82%). In a group of 33 soldiers from the same unit with no signs of the disease IgM-ELISA test was positive in 12 (36.36%) and IgM-IIF in 9 (27.27%). In a group of 40 officers from the unit with the disease outbreak specific IgM antibodies were found by both tests only in the head cook. IgM antibodies were not found in a control group of 102 healthy soldiers from other units of the same garrison. Results of this investigation thermal injury showed variations in immune response to infection in dependence on trauma effect duration (in dependence on the phase). In the early posttraumatic phase organism is capable to react effectively to the inoculated infective agent and in the late posttraumatic phase this reaction is much less effective. Authors conclude that these results may influence upon the thermal injury management strategy in dependence on the trauma effect duration.