Presence Of Mononuclear Cell Infiltrates Research Articles

Nephroprotective effect of Apium graveolens L. against Cisplatin-induced nephrotoxicity

BackgroundCisplatin is extensively used in treating cancers, and its primary side-effect is nephrotoxicity. It accumulates in proximal convoluted tubules where it promotes cellular damage by oxidative stress, apoptosis, and inflammation, etc. In Unani medicine, Tukhm-e-Karafs(Apium graveolens L.) (TK) is mentioned in the literature to manage various kidney ailments due to its diuretic and deobstruent activities. ObjectiveTo investigate the nephroprotective effects of powder of TK in Cisplatin-induced nephrotoxicity in an animal model and to validate the Unani claim of its nephroprotective action. Material and methodsIn curative protocol, cisplatin (5 mg/kg body weight i.p) was administered on day one and powder of TK (500 and 1000 mg/kg p.o.) from the sixth day onwards for ten days. TK (500 and 1000 mg/kg p.o.) was given for ten days and Cisplatin (5 mg/kg body weight i.p) on day 11 in the protective model. At the end of the study, all the animals were sacrificed, and renal biochemical parameters were determined. KIM-1 level was also investigated in the kidney homogenate in conjunction with histopathological inspection of kidney tissues. ResultsSignificant increase in serum creatinine and BUN, presence of mononuclear cell infiltration, tubular dilation and vacuolation in renal histopathology, and increased KIM-1 level confirmed the nephrotoxicity due to Cisplatin. TK's administration protects the kidney as suggested by the changes in biochemical renal function, decreased level of KIM-1, and improvement in histopathological changes. ConclusionThe result advocated that TK prevented renal injury and maintained normal renal function in both models. It may be due to improved clearance of Cisplatin from kidney tubules and reduction in reactive oxygen species (ROS) produced by the inflammatory response.

Hepatic coccidiosis in Serrasalmus rhombeus Linnaeus, 1766 from the Amazon basin: morphological and histopathological aspects.

Calyptospora species are coccids that commonly cause liver infections in fish all around the world. This paper describes the morphology and histopathological characteristics of liver infection caused by Calyptospora sp. in black piranha Serrasalmus rhombeus, from the Capim River, in the municipality of Ipixuna do Pará, state of Pará (Brazil). Specimens were collected, analyzed and necropsied and tissue fragments containing parasites were prepared for histology and scanning electron microscopy. Parasitism was detected in 33.0% of the analyzed specimens, which had spherical oocysts in the liver, with four pyriform sporocysts presenting sporozoites internally. A histological examination revealed oocysts positioned close to blood vessels, causing necrosis and degeneration of hepatic parenchyma, while the presence of mononuclear cell infiltrate and melanomacrophages indicated the onset of an inflammatory process. This is the first record of the genus Calyptospora in fish from the Capim River.

Comparison of Clinic - Histopathologic Findings and Morphometric Measurements of Subclinical Laminitic Claws in Dairy Cattle

The aim of this study was to evaluate the clinic-histopathologic characteristics and to compare the morphometric measurements of healthy and subclinical laminitic claws of dairy cattle at different ages and weights. Non-lame 60 Holstein feet randomly collected from the slaughterhouse were evaluated. The effects of age, body-weight, claw location (right front lateal or right front medial etc), and presence of laminitis were investigated. The claws‘ conformation were evaluated morphometrically with ten measurements (toe length, toe height, outer and inner edges of the claw, heel height, the length of heel, the length of diagonal front wall, dorsal hoof angle, the width and the length of the sole). The claws were classified as normal or laminitic according to the histopathologic findings. The clinical findings of laminitis was confirmed on 71.2% of the claws (n=66). The toe length, toe height, the height of outer and inner edges of the claw, heel height, the length of heel, the length of diagonal front wall were smaller in laminitic claws. The dorsal hoof angle of healthy claws were bigger and statistically significant than the laminitic claws. Small haemorrhagic areas were determined in the parietal corium in the laminitic claws comparing to macroscopically healthy claws. The histopathologic characteristics of the corium of laminitic claws involve the hyperaemia, haemorrhages, oedema, thrombosis of capillaries and presence of mononuclear cell infiltration in dermis, stretching epidermal lamella, necrosis of epithelial cells and detachment of the lamellar basement membrane. According to this study results, contrary to literature, there was not a reliable relation between some changes in morphological structure of the claws and the presence of the laminitis were observed.

Giant Cell Arteritis Diagnosis by Biopsy or Ultrasound: Are we Classifying a Homogeneous Population?

Giant cell arteritis (GCA) is a granulomatous vasculitis with autoimmune origin that is defined by the presence of mononuclear cell infiltrates and the formation of giant cells. It appears in elderly patients and involves the aorta and its branches, particularly the superficial temporal artery. In these patients, rapid diagnosis and immediate initiation of treatment are essential to prevent vascular complications, particularly visual loss and ischemic stroke.

Application of different techniques to detect Toxoplasma gondii in slaughtered sheep for human consumption.

The aim of this study was to investigate occurrence of Toxoplasma gondii in sheep slaughtered in the state of Alagoas, Brazil, by means of different diagnosis techniques. Serum samples and tissues from 100 slaughtered sheep were used. To detect antibodies, the indirect immunofluorescence antibody test (IFAT) was used, and tissues from seropositive animals (cut-off ≥1:64) were submitted to Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC). To assess the concordance between the direct techniques, the kappa test was used. In the IFAT, it was observed that 14% (14/100) of the ovine samples were serum-positive. In the PCR, 21.43% (3/14) of the animals were positive and in IHC, it was observed that 7.14% (1/14) were positively stained for T. gondii in cerebral tissue. Histopathologically, the predominant finding was the presence of mononuclear cell infiltrate in the heart and a perivascular cuff in the cerebrum and cerebellum. The concordance between the direct diagnosis techniques was moderate (k=0.44). Thus, it is important to use different direct techniques in diagnosing toxoplasmosis in naturally infected sheep.

Polymyositis and dermatomyositis as a risk of developing cancer.

Polymyositis (PM) is an idiopathic inflammatory myopathy that affects striated muscles. Dermatomyositis (DM) is an idiopathic inflammatory myopathy with presence of skin symptoms. Both are characterized by acute or subacute onset, symmetrical proximal muscle weakness, the presence of mononuclear cell infiltrates of the muscles and increased activity of muscle enzymes. The treatment still remains glucocorticoids and disease-modifying drugs. Symptoms of PM/DM can be a signal of developing cancer. Known risk factors for cancer in patients with PM/DM are older age, male gender, dysphagia, skin necrosis, cutaneous vasculitis, rapid onset of the disease, elevated creatinine kinase (CK) and C reactive protein (CRP), and an increase in the erythrocyte sedimentation rate (ESR). Recently three new myositis-specific autoantibodies (MSA) predicting the risk of cancer have been discovered: melanoma differentiation-associated protein 5 (anti-MDA-5), transcription intermediary factor 1γ (TIF-1γ), and nuclear matrix protein NXP-2.

English

BACKGROUND AND OBJECTIVES: Alopecia Areata refers to a common usually reversible condition in which patchy loss of hair occurs characterized by presence of mononuclear cell infiltrates developing in and around anagen hair follicles and causing circumscript hair loss. Alopecia Areata has an auto immune pathogenesis mediated by helper T-cells. Since tacrolimus suppresses helper T-cell activity its effect on alopecia areata was studied. METHODS: The study was conducted in the dermatology OPD department in Fr. Muller Medical College Hospital, Mangalore. A total of 50 cases of alopecia areata were taken up for the study. The period of study was from October 2004 - April 2006, the study included 33 males and 17 females. An informed consent was obtained from all the patients. Hair root examination, VDRL, haemogram, Liver Function Test (LFT), Renal Function Test (RFT) and other relevant investigations were done. Focal sepsis was ruled out in all patients. Tacrolimus 0.03% cream was applied once daily and reviewed once in two weeks for twelve weeks. Qualitative data wise score analysis was done by using Friedman's test, Mean, Standard Deviation, Chi-Square test and 'p' value (test of significant) was noted. Response was graded as failure, poor, good and excellent objectively. RESULTS: 6 (12%) patients with scalp involvement had poor response. 20 (40%) patients of alopecia areata on scalp, 5 (10%) patients on face and 2 (4%) patients each on eyebrow and moustache showed good response. 2 (4%) patients with ophiasis pattern also showed good response. 8 (16%) with scalp and 5 (10%) on face had excellent response. None of the patients had any cutaneous or systemic side effects to 0.03% tacrolimus cream during the study period. INTERPRETATION AND CONCLUSION: 10 (20%) patients had associated personal history of atopy and 8 (16%) patients showed exclamatory mark hair. 6 (12%) patients who poorly responded to 0.03% tacrolimus cream had nail pitting. Topical tacrolimus is effective and could be the other modality in treating alopecia areata, however higher strength (0.1%) may give a better result. Being non-irritant topical therapy it could be a treatment modality of choice in children. Absence of cutaneous side effects makes it suitable for use on the face.

Spontaneous poisoning by Ricinus communis (Euphorbiaceae) in cattle

The aim of this study is to report cases of spontaneous poisoning of cattle by Ricinus communis (castor beans) in Paraíba, a semiarid region of northeastern Brazil. The cases were observed in 2 herds on neighboring properties in 2013. Clinical signs developed within 6-24 h and consisted of weakness, tachycardia, dyspnea, profuse watery diarrhea, dehydration, depression, instability, cramps, permanent lateral recumbency and death within 48-72 h. Of the 60 cattle at risk, 19 were affected and 14 died. Five fully recovered after the course of 12 days. Three animals were necropsied. The main gross lesions were hemopericardium, hemothorax, pulmonary edema, petechial hemorrhages in the epicardium and endocardium, ecchymoses at the papillary muscles and suffusions on the intercostal muscles. Hemorrhages were also observed in the abdominal cavity, spleen and mucosa of the abomasum and small intestine. The rumen content was liquid with a large amount of castor bean seeds. There were circular, whitish and focally diffuse areas in the liver parenchyma. The main microscopic lesions consisted of multifocal coagulative myocardial necrosis with the presence of mononuclear cell infiltration and varying degrees of bleeding between cardiac muscle fibers. The abomasum and small intestine mucosae and submucosa had mild edema and mononuclear and polymorphonuclear inflammatory cell infiltration. The diagnosis of R. communis was based on the history of plant consumption, clinical signs, pathology of the disease and the presence of large amounts of castor bean seeds in the forestomachs.

Effects of modified Haizao Yuhu Decoction in experimental autoimmune thyroiditis rats

Modified Haizao Yuhu Decoction (MHYD), based on clinical experience, has been used for approximately 500 years and famous for its efficiency in treating thyroid-related diseases. To investigate therapeutic effects of MHYD on thyroid-related autoantibodies and thyroid hormone in experimental autoimmune thyroiditis (EAT) rats, and to provide evidence for the immunomodulatory potential during antigenmediated immunostimulation. The effect of MHYD was investigated in EAT rats induced by subcutaneous injection of porcine thyroglobulin with Freund's adjuvant (complete and incomplete, CFA and IFA). MHYD was fed to EAT rats daily at a dose of 100mg/kg for 10 weeks. The presence of mononuclear cell infiltration in thyroid tissues were examined to assess the severity of EAT. The levels of serum T3, T4, TgAb and TPOAb were determined by radioimmunoassay and protein expressions of TRAIL were detected by immunohistochemistry. MHYD exhibited a significant protective effects by reversing serum T3, T4, TgAb and TPOAb levels, histopathological changes and TRAIL protein expression of thyroid in EAT model rats. Sargassum fusiforme and Radix Glycyrrhiza in the formulation could be responsible for the immunomodulatory effects of MHYD. These results showed MHYD playing a role as an immunomodulator and TRAIL inhibitor during antigenmediated immunostimulation, may warrant further evaluation as a possible agent for the treatment of Hashimoto's thyroiditis.

Histological analysis of gastritis and Helicobacter pylori infection in patients with early gastric cancer: a case-control study.

Infection with Helicobacter pylori is associated with an increased risk of gastric adenocarcinoma. However, most patients with H. pylori infection will not develop gastric cancer. The aims of the present study were to examine which histological features, including H. pylori infection, would increase the risk of gastric cancer using a case-control study. Three gastric biopsy specimens were taken from 72 patients with early gastric cancer and 72 age- and sex-matched control subjects. The grade of gastritis was examined according to the updated Sydney System. The presence of H. pylori infection was determined by serology and histology. Odds ratio (OR) of developing gastric cancer was calculated for H. pylori positivity and histological features using conditional logistic regression. For patients with H. pylori infection, histological features in cancer patients and control subjects were compared. The OR of the presence of mononuclear cell infiltration in the corpus and intestinal metaplasia in the angulus were significantly elevated. The grade of mononuclear cell infiltration in the corpus and antrum was significantly higher in both types of cancer patients than controls. Glandular atrophy and intestinal metaplasia were increased in patients with intestinal-type cancer in the angulus and antrum. Bacterial density in the corpus and polymorphonuclear cell infiltration in the antrum were increased in patients with diffuse-type cancer. Severe chronic gastritis induced by H. pylori infection seems to be associated with diffuse-type gastric cancer. Glandular atrophy and intestinal metaplasia, which occur in gastric mucosa with chronic inflammation, are significantly associated with intestinal-type cancer.

Altered expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in acutely rejected myocardium and coronary arteriosclerosis in cardiac allografts of nonhuman primates.

Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are important in any process of tissue remodeling. However, there is no report evaluating their expression in cardiac allografts in human or non-human primates. Heterotopic cardiac transplantation was performed on Japanese monkeys. Subjects were treated with chimeric anti-human lymphocyte function-associated antigen-1 monoclonal antibody for 2 weeks. Heart grafts were harvested at days 1-95 (n = 7). Native monkey hearts were used as controls (n = 2). We examined expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 using immunohistochemistry and in situ reverse transcriptase polymerase chain reaction (RT-PCR). In the myocardium, the expression of MMP-2 was increased in the spindle-shaped cells of acutely rejected myocardial interstitium and prior to the presence of mononuclear cell infiltration at days 1-41. TIMP-1 and 2 expression was enhanced in association with the progression of fibrosis at days 40-95. In the coronary arteries of chronically rejected allografts, enhanced MMP and decreased TIMP expression was observed in both thickened intima and media at days 40-95. The medial MMP mRNA expression was observed before the development of intimal thickening occurred at days 7-28. MMPs are critical for the progression of acute and chronic rejection, and TIMP predominance plays important roles in fibrosis in association with acute rejection. Expression of MMPs and TIMPs is a sensitive indicator of acute and chronic cardiac rejection.

Immunological Responses to Injury and Grafting in the Central Nervous System of Nonhuman Primates

Allogeneic transplantation for the therapy of human Parkinson’s disease is being considered as a viable approach at several clinical centers worldwide. As an attempt to understand the basic biology of central nervous system (CNS) transplantation, our laboratory has developed an experimental nonhuman primate model for human Parkinson’s disease and carried out preliminary studies directed at evaluating the potential pathology at the graft site. In addition, studies have been conducted to examine whether such transplantation procedures lead to specific and/or nonspecific immunologic sensitization of the host or results in generalized immunosuppression. Groups of rhesus macaques ( Macaca mulatta) were either controls operated ( n = 6), autografted with adrenal medullary and peripheral nerve tissue ( n = 3), or allografted with fetal mesencephalic tissue ( n= 6). Immunohistological studies demonstrated the presence of mononuclear cell infiltrates as early as 1 wk and up to 1 yr postoperatively, although the frequency of the infiltrating cells declined with time. The infiltrates consisted of variable numbers of cells which express CD2+, CD3+, CD4+, CD8+, CD19+, CD22+, CD25+, and CD68+. There appeared to be no difference in the frequency, kinetics, or phenotype of the infiltrating cells in operative controls compared with recipients of auto- or allografts. Tissue sections obtained postoperatively showed low levels of major histocompatibility complex (MHC) Class I antigens and no detectable level of MHC-Class II antigens in neural tissue. A small aliquot of tissue from the operative site was placed in vitro with media containing interleukin-2 (IL-2), which led to the exudation and growth of mononuclear cells that were predominantly CD4+ cells. Phenotypic studies of peripheral blood mononuclear cells (PBMC) from operative controls, auto- and allograft recipient monkeys performed at varying time periods postoperatively failed to show differences in the frequencies of subsets of T-cells, B-cells, NK-cells, or monocytes. Studies on aliquots of the same PBMC failed to show major functional differences in NK-cells, LAK cells, or response to polyclonal mitogens. Finally, recipients of allogeneic mesencephalic grafts failed to show evidence of donor-specific humoral or cellular sensitization. These data indicate that transplantation of autograft adrenal or allograft fetal mesencephalic tissues in the CNS of nonhuman primate did not induce detectable donor-specific sensitization nor nonspecific immunosuppression.

Immunological responses to injury and grafting in the central nervous system of nonhuman primates.

Allogeneic transplantation for the therapy of human Parkinson's disease is being considered as a viable approach at several clinical centers worldwide. As an attempt to understand the basic biology of central nervous system (CNS) transplantation, our laboratory has developed an experimental nonhuman primate model for human Parkinson's disease and carried out preliminary studies directed at evaluating the potential pathology at the graft site. In addition, studies have been conducted to examine whether such transplantation procedures lead to specific and/or nonspecific immunologic sensitization of the host or results in generalized immunosuppression. Groups of rhesus macaques (Macaca mulatta) were either controls operated (n = 6), autografted with adrenal medullary and peripheral nerve tissue (n = 3), or allografted with fetal mesencephalic tissue (n = 6). Immunohistological studies demonstrated the presence of mononuclear cell infiltrates as early as 1 wk and up to 1 yr postoperatively, although the frequency of the infiltrating cells declined with time. The infiltrates consisted of variable numbers of cells which express CD2+, CD3+, CD4+, CD8+, CD19+, CD22+, CD25+, and CD68+. There appeared to be no difference in the frequency, kinetics, or phenotype of the infiltrating cells in operative controls compared with recipients of auto- or allografts. Tissue sections obtained postoperatively showed low levels of major histocompatibility complex (MHC) Class I antigens and no detectable level of MHC-Class II antigens in neural tissue. A small aliquot of tissue from the operative site was placed in vitro with media containing interleukin-2 (IL-2), which led to the exudation and growth of mononuclear cells that were predominantly CD4+ cells. Phenotypic studies of peripheral blood mononuclear cells (PBMC) from operative controls, auto- and allograft recipient monkeys performed at varying time periods postoperatively failed to show differences in the frequencies of subsets of T-cells, B-cells, NK-cells, or monocytes. Studies on aliquots of the same PBMC failed to show major functional differences in NK-cells, LAK cells, or response to polyclonal mitogens. Finally, recipients of allogeneic mesencephalic grafts failed to show evidence of donor-specific humoral or cellular sensitization. These data indicate that transplantation of autograft adrenal or allograft fetal mesencephalic tissues in the CNS of nonhuman primate did not induce detectable donor-specific sensitization nor nonspecific immunosuppression.

Theiler's Murine Encephalomyelitis Virus (TMEV)-Induced Demyelination: A Model for Human Multiple Sclerosis

Numerous studies suggest that a viral infection, on the appropriate genetic background, may play an important pathogenetic role in the development of multiple sclerosis (MS). Among the several viral models of demyelination that have been investigated during the past two decades, Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease has emerged as one of the best because, similarly to MS, it is based on a combined viral-immune pathogenesis. This review highlights the following salient features of this model. TMEV-induced demyelinating disease is a chronic process, lasting for the life of the animals. Lesions consist of well-demarcated plaques of demyelination, which are strictly related to the presence of mononuclear cell infiltrates. Myelin degeneration is not due to direct viral cytopathic effects, but is rather dependent on the host immune response. Susceptibility/resistance to the disease is genetically regulated, and multiple genes both in and outside the major histocompatibility complex appear to be involved. The best immunological parameter that correlates with susceptibility is the ability of a murine strain to mount a delayed-type hypersensitivity (DTH) response to one or more viral epitopes. The importance of the DTH response against the virus in the pathogenesis of the disease is supported by the prevalent role of TH1 T-helper cells, known to be responsible for DTH responses, in inflamed CNS tissues. The role of DTH responses in the pathogenesis of demyelination is supported by the presence of numerous macrophages in affected CNS and by a direct relationship between the number of macrophages, their persistence in tissues, and the severity of lesions. Macrophages, in addition, are the main reservoir of the virus in the CNS, and their infectability correlates with susceptibility to the disease process. It is hypothesized that following the DTH response to the virus, activated lymphocytes recruit other inflammatory cells, particularly macrophages, into the infected CNS tissues. These nonspecifically recruited cells would secrete a number of proinflammatory molecules and proteases that would destroy myelin as a “bystander effect.”

Lymphocytic traffic and homing into target tissue and the generation of endocrine autoimmunity.

Endocrine autoimmunity is known to be characterized by the presence of specific autoantibodies and from the histopathological point of view by lymphocytic infiltration in the target tissue. The presence of mononuclear cell infiltrates is the pathological hallmark of most endocrine diseases characterized by an autoimmune process directed against antigens expressed on endocrine cells. Infiltrating cells can usually be detected by biopsy or by using other, non-invasive, techniques. However, in endocrine tissue such as the islets of Langerhans and the adrenal glands it is difficult to perform biopsies and diagnosis of the autoimmune process is dependent mainly upon detection of specific autoantibodies. A crucial aspect of endocrine autoimmunity and of all processes of organ specific autoimmunity is why and how lymphocytes migrate from primary lymphoid tissue to their specific targets. This occurs mainly through contact with specific adhesion molecules which enable lymphocytes to adhere to the endothelial vessels in close proximity to the target tissue. In this review we discuss the homing of peripheral mononuclear cells into target endocrine tissues and the mediating role of adhesion molecules.

Detection and localization of cytokine immunoreactivity in retro‐ocular connective tissue in Graves' opthalmopathy

Paracrine interactions between fibroblasts residing in the retro-ocular space and infiltrating lymphocytes/macrophages are thought to be of central importance in the pathogenesis of Graves' ophthalmopathy (GO). Although various roles have been suggested for interferon-gamma (IFN gamma), tumour necrosis factor-alpha (TNF alpha) and interleukin-1 alpha (IL-1 alpha) in GO, their actual presence in Graves' retro-ocular connective tissue has not been demonstrated. We examined surgical specimens obtained during orbital decompression from patients with severe GO (n = 6), and from normal individuals (n = 5), for the presence of IFN gamma, TNF alpha and IL-1 alpha. We used immunohistochemical methods on frozen tissue sections and primary fibroblast cultures, and sodium dodecylsulfate polyacrylamide-gel electrophoresis of tissue extracts and tissue culture supernatants. In addition, immunohistochemical staining of tissues for characterization of the mononuclear cell infiltrates was performed. Aggregates of mononuclear cells in retro-ocular connective and fatty tissue were found in five of six GO tissue specimens, but in none of the control specimens. We detected immunoreactivity for the three cytokines (IFN gamma, TNF alpha and IL-1 alpha) in the five GO tissue specimens that contained mononuclear cell aggregates. In addition, IL-1 alpha immunoreactivity was demonstrable in primary and subsequent GO fibroblast cultures and in their supernatants. In contrast, no immunoreactivity for any of these cytokines was detected in tissue specimens, primary cultures or culture supernatants derived from normal individuals. The presence of mononuclear cell infiltrates and associated immunoreactivity for IFN gamma, TNF alpha, IL-1 alpha in retro-ocular connective tissue derived from patients with GO suggests that the previously demonstrated in vitro functions of these cytokines may indeed be operative in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

Overexpression of class I major histocompatibility complex accompanies insulitis in the non-obese diabetic mouse and is prevented by anti-interferon-? antibody

Overexpression of class I major histocompatibility complex (MHC) proteins on pancreatic islet cells is a characteristic of autoimmune Type 1 (insulin-dependent) diabetes mellitus in humans and in animal models. Studies of post-mortem pancreases from humans with Type 1 diabetes suggest that overexpression of class I MHC proteins may precede mononuclear cell infiltration of the islets (insulitis). Pancreatic histology from the earliest stages of human Type 1 diabetes is rarely available. We have used the non-obese diabetic mouse, given cyclophosphamide to accelerate Beta-cell destruction, to investigate the temporal relationship between the overexpression of class I MHC protein and mRNA and other pathological changes associated with Beta-cell destruction. Prior to cyclophosphamide, immunoperoxidase staining showed that expression of class I MHC proteins was greater on islet cells and infiltrating inflammatory cells of the non-obese diabetic mouse than on islet cells of other mouse strains, whereas staining on exocrine cells was similar. On day three after cyclophosphamide administration, when insulitis had regressed, islet class I MHC protein expression had diminished. A dramatic increase in class I MHC protein expression occurred between days seven and nine, concomitant with reinfiltration of the islets by mononuclear cells; overexpression was seen both on islet cells and on surrounding exocrine cells, but only in the presence of mononuclear cell infiltration. By day 21, class I MHC protein overexpression was again confined to the islets, the exocrine pancreas being free of infiltration. Class I mRNA also increased dramatically by day eight but had virtually returned to normal by day 12.2+ effected by cytokines secreted by activated immuno-inflammatory cells. Class I MHC overexpression should enhance targeting of cytotoxic T cells to Beta cells bearing autoantigen.

Mononuclear cell patterns in the skin. An immunohistological and morphometrical analysis.

A large variety of skin diseases is characterized by the presence of mononuclear cell infiltrates in the dermis and to some extent in the epidermis. We have investigated frozen material of 566 lesions of benign and malignant skin diseases by immunohistological methods with a panel of 20 monoclonal antibodies; quantitative studies using computer-assisted image analysis were additionally performed in 80 specimens. In all reactive lesions, mononuclear cells were arranged in distinct compartments simulating the architecture of normal lymphatic tissue. A qualitatively uniform T cell compartment (mainly helper-inducer T lymphocytes, suppressor-cytotoxic T lymphocytes, Langerhans' cells) with slight quantitative differences was found in all inflammatory skin diseases, in peritumoral infiltrates, and also in normal skin. The B cell compartment (B lymphocytes, few T lymphocytes and monocytes) is only rarely seen (some B cell lymphomas and pseudolymphomas). The monocyte compartment (monocytes, few T lymphocytes) associated with a T cell compartment is typical for granulomatous skin lesions. The epidermis forms a separate functional region, which might evolve into a lymphoepithelial compartment in diseased skin. Low-grade malignant lymphomas of the skin display similar architectural patterns as reactive lesions, whereas high-grade malignant lymphomas do not. Obviously the mononuclear cells in the skin are usually arranged in compartments and form a limited number of distinct patterns. As similar patterns are found in diseases that are completely different with respect to clinical appearance, histological features, cause, and outcome, the patterns are not disease specific but reflect general anatomical-functional relationships of inflammatory cells and the skin.

Clonal analysis and in situ characterization of lymphocytes infiltrating human breast carcinomas.

T lymphocytes were isolated from tumor biopsies in 13 patients with breast carcinomas. Immunohistology with monoclonal antibodies confirmed the presence of mononuclear cell infiltrates composed primarily of T lymphocytes in all tumors studied. While the proportion of T lymphocytes expressing the T4 or the T8 surface marker varied from tumor to tumor as determined by morphometric analysis, T8+ cells were more numerous than T4+ cells in 8/12 breast tumors studied. Relatively few T cells (less than 10% in 11/12 tumors) were in an activated state as judged by the surface expression of HLA-DR antigens or the receptor for interleukin-2 (IL-2). In 1 case 20% of the infiltrating mononuclear cells were expressing the IL-2 receptor. The tumor infiltrating lymphocytes (TIL) recovered from 10 tumors were cloned in a microculture system that permits proliferation of nearly 100% of normal peripheral blood T lymphocytes (PBL-T). In contrast to normal and autologous PBL-T, frequencies of proliferating T lymphocyte precursors (PTL-P) were depressed (less than 0.01) in 7/10 TIL preparations indicating a decreased responsiveness of TIL to phytohemagglutinin at the single-cell level. The frequency of PTL-P was noticeably higher in 2 cases (0.03 and 0.09) and close to normal in 1 case (0.39). A total of 170 clones were expanded in vitro and analyzed for different functional capabilities. Most of these clones expressed the T4+/T8-phenotype (73%) and strikingly 53% of these T4+/T8- clones were cytolytic in a lectin-dependent assay, a functional subset which is uncommon among normal PBL-T. Some clones (10%) lysed allogeneic breast tumor cells (MCF7). Only 15% of the clones displayed natural killer activity. Among the cytolytic clones, 17 of 31 tested were also IL-2 producers irrespective of the T4 or T8 phenotype. Our results show that human mammary carcinomas contain many infiltrating T cells with cytolytic potential. Interestingly, among the proliferating cytolytic T cell clones (56% of the microcultures), many expressed the T4+/T8- phenotype. These findings may indicate that the in situ cytolytic reaction (against unknown antigens) is associated preferentially with class II antigens.

Scleroderma

Systemic scleroderma or progressive systemic sclerosis is a connective-tissue disease characterized by vascular alterations and severe fibrosis. The disease has a predilection for female patients and affects all races and ages, although it is more common during the third and fourth decades of life. The sites most frequently involved are the skin, esophagus, lung, and kidney. Patients with<i>CREST syndrome</i>(calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasias) seem to have a more benign clinical course than those with the diffuse form of the disease, which has a more serious prognosis owing to the high incidence of severe pulmonary fibrosis and kidney insufficiency. The pathophysiology of scleroderma remains poorly understood. Two schools of thought prevail: some believe that the primary event is vascular damage, and others propose a derangement in collagen metabolism as the initial event.¹More recently, emphasis has been placed on the presence of mononuclear cell infiltrates