T Lymphocytes and mononuclear phagocytes in the skin infiltrate of systemic and discoid lupus erythematosus and Jessner's lymphocytic infiltrate

T Lymphocytes and mononuclear phagocytes were measured quantitatively with the histochemical acid α-naphthyl acetate esterase (ANAE) method from paraffin sections of skin affected by systemic and discoid lupus erythematosus and by Jessner's lymphocytic infiltrate. The composition of the patchy cutaneous mononuclear cell infiltrates was similar in these three disorders.     

Hypochlorous acid induced activation of human neutrophil and gingival crevicular fluid collagenase can be inhibited by ascorbate

Abstract— Interstitial collagenase either obtained from human neutrophils by phorbol myristate acetate (PMA) induced degranulation or isolated from human gingival crevicular fluid was found to be activated by addition of an oxidative agent, hypochlorous acid (HOCl). Collagenase released by PMA stimulated neutrophils was completely in latent form but underwent partial autoactivation during 16 h incubation at 22°C in the presence of soy bean trypsin inhibitor. The partial autoactivation was potentiated to complete activation of released collagenase after addition of exogenous HOCl. Ascorbate prevented this activation of neutrophil collagenase. Isolated human gingival crevicular fluid collagenase represented an apparent M_r of 70 kD in completely latent form, whereas 70/54 kD enzyme species were detected for partially autoactive form of the enzyme. Western blot analysis of gingival crevicular fluid using a polyclonal antibody raised against purified human neutrophil collagenase revealed the same 70/54 kD molecular forms of the enzyme. The latent gingival crevicular fluid collagenase was also activated by HOCl and this activation could be prevented by ascorbate. Activation of the 70 kD latent collagenase by HOCl as well as by other non-proteolytic activators such as an organomercurial compound (phenylmercuric chloride) and a gold (I) compound (gold thioglucose) was not associated with detectable changes in apparent M_r, whereas trypsin activation resulted in fragmentation of 70 kD enzyme to 54 kD species. Our results provide further evidence for the neutrophil origin of gingival crevicular fluid collagenase and suggest that, in addition to proteolytic activation, oxidative and antioxidative agents seem to be able to regulate neutrophil collagenase activity.     

The relation of different inflammatory cell types to the various parenchymal components of rejecting kidney allografts

Histochemical and immunoperoxidase techniques were used to characterize the spatial relationships of various inflammatory cell types to the different transplant structures in human renal allograft rejection. T lymphocytes were identified by acid α-naphthyl acetate esterase (ANAE) staining, plasma cells by intracyto-plasmic immunoglobulin, mononuclear phagocytes by intracytoplasmic 'dispersed' ANAE reaction and/or lysozyme staining and granulocytes by intra-cellular lactoferrin. In the two cases of acute rejection the infiltrate around the blood vessels consisted mainly of lymphocytes, whereas the infiltrate around the tubules and within the glomerular tufts consisted mainly of mononuclear phagocytes. In acute rejection only a few plasma cells and granulocytes were seen. In the single case of chronic rejection studied, the lymphocytes were no longer concentrated exclusively around the blood vessels, but diffusely distributed throughout the kidney parenchyma. The different distribution of various inflammatory cells may reflect differences in the functions of these cell types in graft destruction.     

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.     

Inflammatory cells, IgA, C3, fibrin and fibronectin in skin lesions in dermatitis herpetiformis

Skin lesions were produced by application of 50% potassium iodide to twelve patients with dermatitis herpetiformis (DH). Perivascular cellular infiltrates were found to be characteristic of developing lesions. The cells were mainly round cells; α-naphthyl acetate esterase staining revealed that in 24-h lesions the mean percentage of T-lymphocytes was 43%, that of mononuclear phagocytes 6% and that of non-T/non-M cells (mainly B-lymphocytes), 44%. The percentage of the latter was highest (mean 81%) in 6-h specimens, suggesting that these cells are participating in the early stages of lesion formation. The infiltrating cells in dermal papillae and within subepidermal vesicles were predominantly polymorphonuclear leukocytes (mean 86%), with some mononuclear phagocytes and non-T/non-M cells. Immunofiuorescence examination confirmed that fibrin deposition is characteristic of the initial lesions of DH and showed that the same is true of fibronectin. Seven out of eight patients had fibronectin deposits in dermal papillae. IgA was found in all and Ci in most of the specimens and, with the exception of papillary vesicles and blister cavities, tbe intensity of IgA and C3 fluorescence showed no marked alterations during the development of lesions.     

Immune-inflammatory cells in recurrent oral ulcers (ROU)

Abstract— Tissue lesions from eight patients with recurrent oral ulcers (ROU) were subjected to detailed immunohistopathologic studies. In five patients, a specimen of an unaffected area from the opposite site was obtained. The main inflammatory cells in situ were CD3 positive T lymphocytes, with CD4 cells forming approximately half (range 30-60%) and CD8 cells 20% (range 10-30%) of all cells. CD19 positive B lymphocytes formed 5-12% of all cells. Furthermore, 45% (range 15-65%) of all lymphoid cells had signs of previous antigenous contact and had helper/inducer CDw29 type. Suppressor/inducer CD45R cells formed only about 20% (range 7-50%) of all cells. Although this observation suggests involvement of antigen as a causative and/or triggering stimulus, elements of a non-specific inflammatory response were observed as well. Endogenous peroxidase-positive neutrophils were present at the ulcer site, and were occasionally observed intravascularly and in th extracellular matrix in areas characterized by inflammatory mononuclear cell infiltrates. Although the proportion of endogenous peroxidase-positive, recently recruited monocytes was low, CD11b and nonspecific esterase-positive mature tissue macrophages formed about 14% (range 5-35%) of all inflammatory cells in situ, particularly at the periphery of the lymphoid cell infiltrates. Mast cells were also observed in all samples studied, forming 2-5% of inflammatory cells in the richly vascularized connective tissue beneath the basement membrane. In the specimens from clinically unaffected areas, inflammatory cells were rare. Our observations stress the multifaceted nature and participation of multiple effector systems in the local tissue pathogenesis of ROU.     

Proliferating Cells in the Synovial Fluid in Rheumatic Disease

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.     

Allergic and toxic contact dermatitis: inflammatory cell subtypes in epicutaneous test reactions

Histochemical and immunohistochemical techniques were used to identify T lymphocytes, mononuclear phagocytes and plasma cells in situ from allergic and toxic epicutaneous test reactions. Intraccllular α-naphthyl acetate esterase (ANAE), endogenous peroxidase and immunoglobulin were used as markers for inflammatory cells. In allergic contact dermatitis 76 ± 7% of all cells were ANAE-positiveT lymphocytes, 13 ± 6% mononuclear phagocytes and 12 ± 6%), ANAE-negative cells. In toxic skin lesions the corresponding values were 64 ± 20%, 18 ± 15% and 18 ± 6%, respectively. There were no statistically significant differences between the allergic and toxic skin reactions. The basic reaction type in allergic and toxic contact dermatitis seems to be similar, with possibly some qualitative and quantitative differences.