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V Bergroth

Publications and source records attributed to V Bergroth.

At least 55 records · Page 3Linked to original sources
PubMed1986

Expression of MHC class II antigen, interleukin-2 receptor, transferrin receptor and gp 40/80 glycoprotein during different phases of a normal PHA-driven lymphocyte activation in vitro.

This study characterizes the temporal profile of MHC class II antigen, interleukin-2 receptor, transferrin receptor and gp 40/80 glycoprotein lymphocyte activation markers in relation to each other during different phases of PHA-dependent cellular activation in vitro. Binding of these monoclonal lymphocyte activation-probes was visualized by the avidin-biotin-peroxidase complex method. Maximum PHA-dependent MHC class II antigen expression of 27 +/- 3% was observed on culture day 1, but later no significant differences were observed in MHC class II antigen expression between PHA-driven or culture media-containing control cultures. On the contrary, interleukin-2 receptor (78 +/- 6%) and transferrin receptor (75 +/- 5%) expression reached a maximum on culture day 3, coinciding with a maximum proliferative response. On culture day 5, when 3H-thymidine incorporation was already on the decline, gp 40/80 glycoprotein reached a maximum PHA-dependent expression of 78 +/- 2%, which differed significantly from interleukin-2 receptor (60 +/- 8%, p less than 0.05) and transferrin receptor (51 +/- 8%, p less than 0.01) expression. This study suggests that MHC class II antigen, interleukin-2 receptor, transferrin receptor and gp 40/80 glycoprotein, although all of them are lymphocyte-activation markers, differ as to the chronological sequence of their appearance and disappearance. Their combined use in lymphocyte-activation marker profile assay therefore gives valuable information about the lymphocyte activation state.

Antigens, Surface↗
PubMed1986

Phenotypic in situ characterization of lymphocytes in mild alcoholic liver disease.

The subsets and expression of activation markers of inflammatory mononuclear cells in mild alcoholic liver disease were studied using monoclonal antibodies and an avidin-biotin-peroxidase complex (ABC) method. There were few B cells and monocytes in situ. Most of the mononuclear cells were of T cell origin, T4 and T8 positive cells being evenly distributed in a ratio of 2 to 1. This contrasts with the present and earlier findings on autoimmune liver diseases. The frequency of Ia (HLA-DR) positive cells varied between 15 and 30% of all mononuclear cells, which indicates activation of the local T cells. This implies that T cells in alcoholic fatty liver are not innocent bystanders but actively participate in the local inflammatory process. Staining for Tac, 4F2 and T9 was completely negative, however. These findings suggest that the local T cell activation may be incomplete and that it is not as extensive as in autoimmune liver diseases.

Humans↗
PubMed1986

Focus score in the diagnosis of Sjögren's syndrome.

According to current agreements, the verification of SS should consider all three components of diagnostic relevance: focal sialadenitis, keratoconjunctivitis sicca and an associated disease. Focus score (the number of mononuclear cell infiltrates containing at least 50 inflammatory cells in a 4 mm2 glandular section) of greater than 1 is probably the single most important test result in the diagnosis of the oral component in SS. Without such quantitation mild, nonspecific inflammation is often falsely classified as focal sialadenitis suggesting an underlying SS. This was confirmed by a retrospective study of a slide library, which showed that most cases descriptively diagnosed as focal sialadenitis actually had focus scores of less than 1. Furthermore, conditions other than SS can result in focus scores of greater than 1. Firstly, in 8/17 patients, sialolithiasis of the submandibular glands was associated with focus scores of greater than 1, usually with little or no histological signs of obstruction and/or infection. Secondly, focus score values of greater than 1 were found in the labial salivary glands in 6/40 coroner's autopsies although the subjects had no clinical history or findings suggesting SS; simultaneous specific involvement of both salivary and lacrimal glands was, however, observed in 0/40 of the subjects. Based on our observations we suggest that a postmortem diagnosis of SS should be made on the demonstration of focus score of greater than 1 in both the labial salivary glands and the lacrimal glands.

Humans↗
PubMed1985

Proliferating cells in the synovial fluid in rheumatic disease. An analysis with autoradiography-immunoperoxidase double staining.

The subtype of the proliferating cells in the synovial fluid of patients with rheumatoid arthritis (RA), ankylopoietic spondylarthrosis (SPA), and osteoarthritis (OA) was studied with autoradiography-immunoperoxidase double staining. Of all spontaneously proliferating synovial fluid cells in chronic arthritis, 59 +/- 4% displayed T8 differentiation marker, whereas T4 (21 +/- 4%) and B (2 +/- 1%) cells were few. Of all T4+ and all T8+ lymphocytes, 0.55 +/- 0.1% and 0.90 +/- 0.1%, respectively, incorporated [3H]thymidine. The [3H]thymidine labelling index for B cells was 0.30 +/- 0.1%. This was in contrast to OA, in which no proliferating lymphocytes were observed in the synovial fluid. Our findings suggest that the predominance of proliferating T8+ cells in the synovial fluid reflects an underlying chronic inflammation. Because RA and SPA synovium is a site of intense immunoglobulin production, our finding of the predominance of activated, proliferating T8+ cells may also reflect a dissociation between phenotype and function as a reason for the chronicity of the joint inflammation.

Adult↗
PubMed1985

Lymphocyte activation in rheumatoid arthritis synovial fluid in vivo.

Monoclonal antibodies were used in avidin-biotin-peroxidase complex staining for activation marker analysis of rheumatoid synovial fluid cells. Although Ia expression indicates T cell activation, cells displaying receptors for interleukin 2 (Tac)-and transferrin receptor (T9)- positive proliferating cells were relatively few. Similarly, activated terminal effector cells of suppressor/cytotoxic nature were scarce in rheumatoid synovial fluid, as suggested by a low expression of Tac and 4F2 markers. The in vivo situation in the rheumatoid arthritic (RA) joint does not seem to be due to the inability of synovial fluid lymphocytes to become activated, because mitogen stimulation in vitro, in spite of a low proliferative response, induced expression of all the activation markers studied. The relevance of the present observations to the down-regulation of the active, inflammatory-immune response in situ is speculative, but the data show that in spite of T-cell activation and Ia expression, activated terminal effector cells of suppressor/cytotoxic nature are few in the RA joint in vivo.

Antibodies, Monoclonal↗
PubMed1985

Synovial fluid cells in Reiter's syndrome.

Synovial fluid cells in Reiter's syndrome were studied by cell subset specific monoclonal antibodies and avidin-biotin-peroxidase complex staining. Mean leucocyte count was 9842/mm3 (9.842 X 10(9)/l), and 71% of all cells were polymorphonuclear leucocytes. 26 +/- 11 (SEM)% and 47 +/- 5% of all mononuclear cells in synovial fluid were M1+ monocytes and Ia+ cells, respectively. T11+ T lymphocyte was the predominant synovial fluid mononuclear cell (61 +/- 8%) but, in contrast to the inflammatory joint effusions in rheumatoid arthritis, T4+ cells clearly outnumbered T8+ cells in Reiter's syndrome. Thus the synovial fluid in Reiter's syndrome contains the immunocompetent and accessory cells required for immune response, which in fact is activated as suggested by lymphocyte Ia expression. Furthermore, in contrast with rheumatoid arthritis inducer/helper cells with T4 phenotype seem to be involved preferentially in the local pathogenetic mechanisms in Reiter's syndrome.

Adult↗
PubMed1985

Cellular immunohistopathology of acute, subacute, and chronic synovitis in rheumatoid arthritis.

Cellular inflammation in rheumatoid arthritis (RA) synovial membrane was studied in biopsy specimens taken at different stages of synovitis and disease. Patients were classified into three subgroups: acute RA, subacute RA, and chronic RA. Inflammatory cells were characterised by a histochemical esterase method and immunohistochemical peroxidase-antiperoxidase (PAP) and avidin-biotin-peroxidase (ABC) staining. The amounts and distribution of inflammatory cells were different in various stages of the synovitis. In acute onset RA monocytes and granulocytes predominated, suggesting that the beginning of rheumatoid inflammation is similar to inflammatory reaction in general. The presence of T cells and also of plasma cells in subacute RA suggests underlying subclinical changes also in apparently healthy joints in RA. The most typical feature of prolonged synovitis in chronic RA was its intensity, characterised by the presence of large T cell and plasma cell infiltrates. Our findings suggest that the immunological mechanisms are secondary to the tissue damage caused by the initial inflammatory events of unknown cause. However, the immunological mechanisms may still play a central role in the aetiopathogenesis, because findings in chronic RA suggest a defective down-regulation of the immune response.

Acute Disease↗
PubMed1985

Langerhans cells in SLE skin. A role in lymphocyte migration and activation in situ.

Langerhans cells in SLE skin and mucosa were studied by using monoclonal anti-T6 and anti-Ia antibodies and avidin-biotin-peroxidase complex staining. Langerhans cells were present in cell infiltrates of natural SLE skin lesions. In developing early skin lesions induced with test antigen, Langerhans cells were the predominant inflammatory cells, suggesting that migration of Langerhans cells into the inflammatory site in fact precedes lymphocyte migration. These findings agree with the hypothesis of the role of dendritic cells in lymphocyte circulation and antigen presentation under physiological and pathological conditions. Many of the infiltrating local T lymphocytes in both natural and test antigen induced SLE lesions were activated Ia+ cells. This suggests an active immune response in both natural and test antigen induced skin lesions in SLE. The low immunomodulatory T4/T8 ratio in situ in SLE skin lesions might suggest that T8+ cells exert immunosuppressive control on the local inflammation.

Cell Movement↗
PubMed1983

Lymphocyte infiltrations of the gastric mucosa in Sjögren's syndrome. An immunoperoxidase study using monoclonal antibodies in the avidin-biotin-peroxidase method.

Biopsy specimens of the gastric mucosa from 7 patients with primary or secondary Sjögren's syndrome were studied using the immunohistochemical avidin-biotin-peroxidase complex (ABC) method. Monoclonal antibodies (OKT series) were used as a primary layer to detect the surface antigens of various lymphocyte subsets in situ. Chronic inflammation with mononuclear cell infiltrates and/or glandular atrophy was seen in all 7 biopsy specimens. Immunohistochemical staining showed that the cell infiltrates consisted mainly of OKT3-positive T lymphocytes. In 1 patient plasma cells predominated at the sites of inflammation. Most of the T cells were OKT4-reactive lymphocytes, the OKT4/OKT8 ratio varying between 3 and 7. These findings from the gastric mucosa are in agreement with those obtained from the salivary glands of patients with Sjögren's syndrome and confirm the view that Sjögren's syndrome is a systemic disease affecting many organs, rather than a local disease restricted only to some exocrine glands.

Antibodies, Monoclonal↗
PubMed1983

Comparison of various immunohistochemical methods. Demonstration of extracellular antigens in cryostat sections.

Immunofluorescence (IF) and immunoperoxidase (IP) methods for the demonstration of IgG bound to intercellular substance in pemphigus skin were compared. Direct and indirect IF and IP methods had comparable sensitivities. The use of undiluted primary antiserum gave false negative results with the indirect methods. This was probably due to the prozone effect. The higher sensitivity of multiple layer techniques permitted the use of lower titers of the primary antiserum and gave positive staining results even in tissue fixed under less than optimal conditions. Enzyme bridge (EB) and peroxidase-antiperoxidase (PAP) methods gave more dermal background staining than the avidin-biotin-peroxidase complex (ABC) method. In addition, the staining results obtained with the EB and PAP methods were dependent on optimum dilution of different antiserum layers against each other. False negative staining was seen as a result of the secondary antiserum being too dilute in relation to the primary antiserum. This was probably because both Fab parts of the link antibody were bound to the primary antiserum. The ABC method was less sensitive in this respect. With all methods, fixation for 5 min at +4 degrees C in acetone gave the best preservation of immunoreactive determinants of IgG. The results indicate that when an immunohistochemical method is chosen for the demonstration of extracellular antigens in cryostat sections, the sensitivity of the method and preservation of the immunoreactive determinants in the sections, the time needed for staining, access to a fluorescent microscope and reproducibility should be taken into consideration.

Fluorescent Antibody Technique↗
PubMed1983

Cellular infiltrate in situ and response kinetics of human intradermal and epicutaneous tuberculin reactions.

Monoclonal antibodies used in the avidin-biotin-peroxidase complex (ABC) method and a histochemical azo-dye method for acid alpha-naphthyl acetate esterase (ANAE) were used to identify T lymphocytes and their functional subpopulations, B cells, and mononuclear phagocytes in tuberculin-test reactions. At 6 and 24 hr up to 52% of all dermal inflammatory cells in situ were T6+, whereas at 72 hr no T6+ cells were observed in the dermis. This suggests that in the initial phases of tuberculin reactions the epidermal Langerhans cells are mobilized to the perivascular spaces in the dermis. At 6, 24, and 48 hr T3+ T lymphocytes usually formed the main inflammatory cell type in situ. The high proportion of Ia+ T lymphocytes and the variations in the local proportion and numbers of T4+ and T8+ cells suggest that T cells play an active role in the generation of a positive tuberculin reaction. At 72 hr T-pattern ANAE+ lymphocytes accounted for 64 +/- 9 and 35 +/- 10% of all cells in situ in epicutaneous and intradermal tuberculin reactions, respectively (P less than 0.05). The corresponding values for M-pattern ANAE+ macrophages were 27 +/- 9 and 53 +/- 10% (P less than 0.10). This indicates that the recruitment of mononuclear phagocytes is delayed if tuberculin is applied epicutaneously rather than injected intradermally.

Humans↗
PubMed1983

Immunocompetent cells in labial salivary glands in secondary Sjögren's syndrome associated with SLE.

T and B lymphocyte (sub)populations were identified by monoclonal hybridoma antibodies (the avidin-biotin-peroxidase complex method), in the periductal lymphocyte-rich infiltrates in the labial salivary glands of 8 patients with secondary Sjögren's syndrome (2 degrees SS) associated with systemic lupus erythematosus (SLE). 59 +/- 7% and 17 +/- 3% of inflammatory round cells in situ were T3-positive and surface(SIg)- or cytoplasmic(CIg) immunoglobulin-positive, respectively. This suggests a local T lymphocyte dominance in salivary glands in 2 degrees SS associated with SLE. The local ratio of cells expressing T inducer/helper: T suppressor/cytotoxic phenotype was 3.5 +/- 0.8 (range 0.9-7.6) indicating large variations between individual patients. 46 +/- 9% of all inflammatory cells in situ were endogenous peroxidase-negative, Ia-positive cells, suggesting an active role for the locally accumulated T lymphocytes.

Adult↗
PubMed1983

A method for the identification of human peripheral blood T lymphocytes by sequential immunogold and esterase double staining.

A double staining method involving the sequential use of monoclonal OKT hybridoma antibodies applied in the colloidal immunogold method and followed by a simultaneously capturing azo dye method for the detection of acid alpha-naphthyl acetate esterase (ANAE) is described. Mononuclear leukocytes isolated from human peripheral blood using a Ficoll-Hypaque density gradient were stained. M-pattern ANAE-positive monocytes (diffuse staining) were excluded from the lymphocyte counts. 80 +/- 5% of all lymphocytes were T-pattern ANAE positive (dot-like staining) and 77 +/- 3% were OKT3 positive. 86 +/- 6% of all ANAE-positive T-pattern lymphocytes were also OKT3 positive, and 89 +/- 6% of all OKT3-positive lymphocytes were also ANAE positive. This indicates that ANAE is a good marker for total human T lymphocytes. 53 +/- 10% of human peripheral blood lymphocytes were OKT4 positive and 87 +/- 8% of all OKT4-positive lymphocytes were also ANAE positive. 30 +/- 6% of all lymphocytes were OKT8 positive, and only 26 +/- 18% of all OKT8-positive lymphocytes were ANAE negative. This indicates that ANAE cannot be used to distinguish T-helper and T-suppressor lymphocytes as identified by monoclonal antibodies.

Antibodies, Monoclonal↗
PubMed1983

Factors affecting the immunoperoxidase demonstration of intracellular immunoglobulins and J chain from cytocentrifuged cell smears.

Immunohistochemical methods were used to study 1) the optimum fixation conditions for the preservation of human J chain and immunoglobulin (Ig) immunoreactivity and 2) the relation of J chain synthesis by plasmablasts and plasma cells to Ig synthesis in cell smears of cultured human peripheral blood lymphocytes stimulated with pokeweed mitogen (PWM). J chain was demonstrated using the indirect immunoperoxidase method, and intracellular Ig was demonstrated with the unlabeled antibody--enzyme method. In the sequential double staining procedure, J chain was demonstrated using the indirect immunoperoxidase method followed by the demonstration of Ig with the direct immunofluorescence method. Optimum preservation of J chain immunoreactivity was obtained with fixation in neutral buffered formalin at 22 degrees C for 5 min followed by immediate immunoperoxidase staining. False negative results were seen when the slides were stained 2 weeks after fixation. In PWM-stimulated smears, J chain appeared on day three, simultaneously with or after the onset of Ig synthesis. In double stained smears most IgG-positive cells also showed immunoreactivity for J chain from the third day on.

Fluorescent Antibody Technique↗
PubMed1982

Fixation-dependent cytoplasmic false-positive staining with an immunoperoxidase method.

Fixation-dependent nonspecific staining with the unlabeled immunoperoxidase (PAP) method was studied using paraffin-embedded human spleen sections fixed in various fixatives; the specific primary antiserum was omitted or nonimmunized normal rabbit serum was used. Strong cytoplasmic staining of polymorphonuclear leucocytes and macrophages was found after fixation in acetone, alcoholic formalin (94% alcohol) and absolute ethanol. This staining was mainly produced by the second layer of the PAP method. The most probable explanation of this phenomenon is nonspecific protein-immunoglobulin interaction as a result of alcoholic or acetone fixation of the sections. The present findings point to the importance of controls for each case under study to avoid false-positive interpretations.

Humans↗
PubMed1982

Human ferritin: effects of antigen source and fixation on leucocyte staining by immunoperoxidase technique.

The localization of ferritin was studied in peripheral blood cells and variously fixed tissues with the antibodies against ferritins isolated from human heart and spleen. The unlabelled antibody enzyme method (PAP) was used to detect the binding sites of antibodies. In peripheral blood cell smears both antisera gave rise to strong staining of polymorphonuclear (PMN) cell cytoplasm, whereas the monocytes stained relatively weakly. There were no staining differences between the two antisera. In human spleen sections the spleen ferritin antiserum stained the PMN cells and sinusoidal lining cells, whereas the heart ferritin antiserum stained only PMN cells. Neither of the two antisera stained monocytes in the spleen sections. This finding was observed in specimens fixed in Bouin's fixative, Baker's fixative and neutral formalin. However, the immunoreactivity of ferritin was totally destroyed by some other fixatives (Carnoy's fixative, formol sucrose and glutaraldehyde). These results suggest that ferritin is more readily released from monocytes than from PMN cells, and that mature spleen macrophages contain antigenic determinants of ferritin that are recognized only by anti-spleen ferritin antiserum.

Cytoplasm↗