Demonstration Of Immunoglobulin Research Articles

A RARE CASE OF PLEURODYNIA AFTER THE COVID-19 VACCINE

A RARE CASE OF PLEURODYNIA AFTER THE COVID-19 VACCINE

HISTOPATHOLOGICAL AND IMMUNOFLUORESCENT STUDIES IN CERTAIN SKIN DISORDERS

Background: This study was an attempt to evaluate the role of direct immunofluorescent technique to demonstrate the immunoglobulins in certain skin disorders, which are likely to have immunological mechanism in their pathogenesis.
 Methods: This study was carried out in department of Pathology, Sardar Patel Medical College & Associated group of Hospitals, Bikaner. This study was hospital based study on skin biopsy specimen recieved in the department of Pathology during the study period.
 Results: On immunofluorescent studies of these skin biopsies, 12 out of 16 cases of pemphigus were positive for fluorescence was intracellular area in epidermis. In lichen planus 5 out of 7 cases were positive for fluorescence and commonest site was dermo-epideremal junction. In dermatis-herpatiformis 2 out of 3cases were positive for fluorescence and commonest site was dermo-epideremal junction. IgG was commonest type of immunoglobulin’s demonstrate in 12 out of 16 cases of pemphigus followed by IgM (5 cases), IgA(1cases). In lichen planus IgM was commonest type of immunoglobulin’s demonstrate in 5 out of 7 cases of pemphigus followed by IgG (3 cases), IgA(1cases). In dermatitis herpatiformis IgA was commonest type of immunoglobulin’s demonstrate in 2 out of 3 cases of pemphigus followed by IgM (1 cases).
 Conclusion: We conclude that the demonstration of immunoglobulins in skin biopsies by direct fluorescent technique is a quite useful adjunct in diagnostic confirmation of pemphigus, lichen planus and dermatitis herpetiformis.
 Keywords: Direct immunofluorescence, Pemphigus, Histopathology.

Demonstration of immunoglobulin G antibodies against Onchocerca volvulus excretory-secretory antigens in different forms and stages of onchocerciasis

The excretory-secretory (E-S) products of helminths are considered to comprise immunogenic molecules of high diagnostic value. In the present study, the serodiagnostic potential of the E-S products released in vitro by cultured female Onchocerca volvulus was investigated by enzyme-linked immunosorbent assay ( elisa) and Western blotting using 190 serum samples from persons infected with O. volvulus and unexposed persons. The sensitivity of detection of anti- O. volvulus E-S antibodies was 94% for sera from patients with the generalized form of onchocerciasis and 100% for sera from patients with the chronic hyperreactive form (sowda). 95% of the sera from amicrofilaridermic persons, who subsequently became microfilaridermic within 2 years, reacted with O. volvulus E-S antigens and the donors were therefore regarded as having had a prepatent infection when first examined. These sera gave higher ( P < 0·05) elisa optical densities than sera from the same persons obtained when they had become patent, indicating a loss of antibody reactivity after emergence of microfilariae. The specificity of the E-S elisa was 100% when sera of subjects infected with Wuchereria bancrofti were used, and at least 88% for Mansonella perstans sera. In Western blot analysis, the sera of persons with generalized onchocerciasis recognized 7 protein bands. Many E-S proteins were stained less intensely by the sera of subjects with generalized onchocerciasis than by the sera of sowda patients. Similar antigen bands were demonstrated using sera from the persons with prepatent infections.

Demonstration of immunoglobulin G with affinity for dopamine in cerebrospinal fluid from psychotic patients

Using an enzyme-linked immunosorbent assay, significantly raised concentrations of immunoglobulin G with affinity for the neurotransmitter dopamine were demonstrated in cerebrospinal fluid psychotic patients. We have varied the antigen presentation in order to find a conjugate with low unspecific binding. The conjugation of dopamine to carbodiimide-activated poly- L-glutamic acid and that to activated succinimide ester of biotin are described. The use of glutaraldehyde conjugation is not recommended because of the risk of formation of tetrahydroisoquinolines. A strong correlation ( r = 0.94, P < 0.001) between the results obtained with dopamine conjugated to poly- L-glutamic acid and dopamine conjugated to biotin was observed. Forty-two human cerebrospinal fluid samples from 20

Immunoglobulin Gene Rearrangement in Abnormal Lymph Node Hyperplasia

The histologic designation "abnormal lymphoid hyperplasia" is applied to lymph nodes demonstrating varying degrees of architectural effacement and/or cytologic atypia. Although some of these cases may be suggestive of non-Hodgkin's lymphoma, a definitive diagnosis is not possible despite careful morphologic and immunophenotypic studies. Because the demonstration of immunoglobulin and T-cell receptor gene rearrangements by Southern blot analysis provides a sensitive marker of lineage and clonality in lymphoid malignant conditions, the frequency with which such gene rearrangements could be identified in abnormal hyperplasia and their significance were studied. DNA samples from lymph node biopsy samples of 11 patients with abnormal lymphoid hyperplasia were analyzed for rearrangements of immunoglobulin and T-cell receptor genes by Southern blot hybridization. Six of these patients had monoclonal B-cell populations identified by immunoglobulin gene rearrangements; all were found subsequently to have non-Hodgkin's lymphoma by repeated biopsy from 8 days to 46 months later. Two patients with negative Southern blot studies also developed lymphoma, one a T-cell non-Hodgkin's lymphoma and one a cutaneous B-cell non-Hodgkin's lymphoma. Three patients without detectable gene rearrangements showed no evidence of malignant lymphoma at 36-, 45-, and 60-month follow-up evaluations. Southern blot analysis thus identified monoclonal B-cell lymphoid populations in a subset of patients with abnormal lymphoid hyperplasia; the presence of clonal immunoglobulin gene rearrangement predicted progression to overt non-Hodgkin's lymphoma.

Gene Rearrangements in the Diagnosis of Lymphoma/Leukemia: Guidelines for Use Based on a Multiinstitutional Study

The demonstration of immunoglobulin or T-cell receptor gene rearrangements in human lymphoproliferative processes with the use of DNA hybridization has gained great popularity as a sensitive laboratory adjunct to diagnostic hematopathology. The fact that nearly all B- or T-cell malignant lymphomas and leukemias have one or more rearranged antigen receptor genes provides a biologic basis for a diagnostic test. To formally analyze the sensitivity, specificity, and reproducibility of gene rearrangements in the diagnosis of human lymphoproliferative disease, the authors conducted a large, multiinstitutional study. Through a blinded, controlled approach, gene rearrangement analysis of 275 cases was shown to carry a high correlation with conventional phenotyping and histologic diagnosis, with only minor false-positive and false-negative rates. Significantly, no rearrangements were detected in normal lymphoid tissues or carcinomas, sarcomas, or melanomas. In a randomized study of 50 cases, laboratory results showed a high rate of interlaboratory agreement, regardless of the level of previous experience. Furthermore, the reproducibility of interpretation of data (Southern blot autoradiograms) of 192 cases showed high concordance among 11 observers from multiple laboratories. Based on these findings, the authors propose a set of guidelines for interpretation of gene rearrangement analysis that, if carefully followed, renders this a highly reproducible, safe, and accurate addition to the diagnostic regimen for human lymphoproliferative processes.

Temporal Artery Biopsies: Correlation of Light Microscopy and Immunofluorescence Microscopy

Immunopathologic studies are done routinely on biopsy specimens from tissues affected by many autoimmune diseases. To evaluate the role of direct immunofluorescence microscopy (DIFM) in identifying temporal arteritis, the authors reviewed all temporal artery biopsies done over a 30-month period (100 consecutive biopsies). The DIFM, using antibodies to IgG, IgM, IgA, complement, and fibrinogen, had a diagnostic sensitivity rate of 93% and a specificity rate of 87% compared with light microscopy. In biopsy specimens showing arteritis by light microscopy, IgG was demonstrated by DIFM in 85% of cases, IgM in 69%, and IgA in 15%. In one patient, a DIFM staining pattern highly suspicious of temporal arteritis identified a patient with features of clinical temporal arteritis despite negative findings by light microscopy. The demonstration of immunoglobulin by DIFM supports the possible role of humoral immunity in the pathogenesis of temporal arteritis.

(11) Oesophageal strictures in cicatricial pemphigoid

Cicatricial pemphigoid is a rare disease affecting the skin and mucous membranes. It is a disorder characterized by sub-epidermal blisters which heal with fibrosis and scarring. The diagnosis is confirmed by demonstration of immunoglobulins along the basement membrane of perilesional tissue using immunofluorescence. Ten patients with cicatricial pemphigoid who complained of dysphagia were investigated by barium swallow. An upper oesophageal stricture was demonstrated in eight of these. In the remaining two patients indirect laryngoscopy showed hypopharyngeal ulcers which could account for the dysphagia. A benign stricture of the upper third of the oesophagus is a rare finding and cicatricial pemphigoid should be recognized as a possible cause. The other clinical manifestations of cicatricial pemphigoid may be subtle. However, they can be seen on careful examination of the skin and mucous membranes which will enable the diagnosis to be made.

A Technique for the Demonstration of Immunoglobulins in the Cumulus oophorus-Oocyte Complex of the Pig

T r a v n i k P., D. H 0 r k y: A Technique for the Oemons.tration of Immunoglobulins in the Cumulus oophorus-Oocyte Complex of the Pig. Acta vet. Brno, 56, 1987: 487-493. An optimized procedure for the demonstration of immunoglobu­ lins in the pig cumulus oophorus-oocyte complex in semi-thin sections is described. Non-specific porcine immunoglobulIns of class G were demonstrated by means of peroxidase~label led goat antiserum, and specific immunoglobulins of class G against Aujeszky's disease virus were demonstrated using the correspond­ ing antigen and peroxidase-labelled porcine antibodies against this antigen. A most convenient mode of removing the embedding medium was developed, the composition of the incubation solution for the demonstration of peroxidase activity was improved and a number of checks on th~ specificity of the reaction were tested. The procedure proved most useful for the demonstration of specific and non-specific immunoglobulins both in docytes and in the cu­ mulus oophorus. Oocyte, cumulus oophorus, immunoglobulins, immunocytochemistry, semi-thin sections. Light microscopic examination of semi-thin sections provides sufficient information on the location of a substance as to whet­ her it is present in the cumulus·oophorus cells, intercellular spaces of the cumulus, in the zona pellucida, perivitelline space, cytoplasm or in the nucleus of the oocyte. Moreover, semi-thin sections of the cumulus-oocyte complex are easier to prepare than traditional paraffin-embedded sections. In the light of these considerations we chose the immunocytochemical demonstration of specific and non-specific immunoglobulins in semi-thin sections

Demonstration of immunoglobulin M class antibodies to Toxoplasma gondii antigenic component p35000 by enzyme-linked antigen immunosorbent assay.

On the basis that 89% of 48 acute-phase toxoplasmosis patients showed immunoglobulin M (IgM) class antibodies to the 35,000-molecular-weight antigenic component (p35000) of Toxoplasma gondii, as demonstrated by IgM immunoblotting, the antigen was purified by sucrose gradient centrifugation and enzyme labeled for use in an enzyme-linked antigen immunosorbent assay (ELA) for the demonstration of IgM class antibodies to the p35000 component. The ELA showed a specificity of 96% with 139 serum specimens at a serum dilution of only 1:5. The test serologically detected 73 symptomatic acute-phase toxoplasmosis patients; 64 were positive in the 19S IgM indirect immunofluorescent-antibody test, and 9 were negative, although they showed IgM antibodies to p35000, as demonstrated by IgM immunoblotting. Also, the ELA turned out to be independent of IgM rheumatoid factors in six acute-phase toxoplasmosis serum specimens.

Separation of lymphoid and myeloid blasts in the mixed blast crisis of chronic myelogenous leukemia: no evidence for Ig gene rearrangement in CALLA-positive blasts

Recent studies suggest that lymphoid blast crisis cells of chronic myelogenous leukemia (CML) expressing the common acute lymphoblastic leukemia antigen (CALLA) are B precursor cells, based on the demonstration of immunoglobulin (Ig) gene rearrangement similar to common acute lymphocytic leukemia. There is little evidence to suggest whether the cells with similar lymphoid characteristics in the mixed blast crisis of CML are also committed to B cell lineage. A patient in “mixed” blast crisis of CML was studied. On the basis of morphology, cytochemistry, and immunological studies, the blasts were classified as having either lymphoid or myeloid characteristics. A proportion of the leukemic blasts expressed CALLA, whereas others expressed My7 antigen. In order to characterize both populations of cell further, CALLA+ blasts and My7+ (myeloid) blasts were isolated by fluorescence- activated cell sorting. The My7+ cells were highly proliferative in cell culture blast colony assays, retained the Ph1 chromosome, and were indistinguishable from acute myelogenous leukemia blasts. The CALLA+ cells were also Ph1-chromosome positive, but in contrast, were poorly proliferative in vitro. Of particular note was their retention of germline configuration of Ig genes, thus distinguishing them from blasts in the lymphoid crisis of CML. We conclude that the lymphoid component in mixed blast crisis may represent a stage of differentiation prior to commitment to B lineage.

Separation of Lymphoid and Myeloid Blasts in the Mixed Blast Crisis of Chronic Myelogenous Leukemia: No Evidence for Ig Gene Rearrangement in CALLA-Positive Blasts

Recent studies suggest that lymphoid blast crisis cells of chronic myelogenous leukemia (CML) expressing the common acute lymphoblastic leukemia antigen (CALLA) are B precursor cells, based on the demonstration of immunoglobulin (Ig) gene rearrangement similar to common acute lymphocytic leukemia. There is little evidence to suggest whether the cells with similar lymphoid characteristics in the mixed blast crisis of CML are also committed to B cell lineage. A patient in "mixed" blast crisis of CML was studied. On the basis of morphology, cytochemistry, and immunological studies, the blasts were classified as having either lymphoid or myeloid characteristics. A proportion of the leukemic blasts expressed CALLA, whereas others expressed My7 antigen. In order to characterize both populations of cell further, CALLA+ blasts and My7+ (myeloid) blasts were isolated by fluorescence-activated cell sorting. The My7+ cells were highly proliferative in cell culture blast colony assays, retained the Ph1 chromosome, and were indistinguishable from acute myelogenous leukemia blasts. The CALLA+ cells were also Ph1-chromosome positive, but in contrast, were poorly proliferative in vitro. Of particular note was their retention of germline configuration of Ig genes, thus distinguishing them from blasts in the lymphoid crisis of CML. We conclude that the lymphoid component in mixed blast crisis may represent a stage of differentiation prior to commitment to B lineage.

Different fixative-resin combinations and the immunocytochemical demonstration of immunoglobulins (kappa light chains) in semi-thin sections.

The effect of six different epoxy resins on the immunostaining of kappa light chains in tonsilar tissue which had been fixed by one of three fixing variants is described. The results show that not only may a specific resin influence the reaction but this may be enhanced by adopting a particular fixative-resin combination.

Demonstration of immunoglobulin in brains of ferrets inoculated with an SSPE strain of measles virus: use of protein A conjugated to horseradish peroxidase.

Measles virus-specific immunoglobulin G (IgG) has been found in the brains of patients with subacute sclerosing panencephalitis (SSPE), a slowly progressing central nervous system (CNS) disease affecting children. IgG/albumin ratios indicate that the antibodies are probably synthesized in the CNS. In a ferret model system, protein A conjugated to horseradish peroxidase (PrAPx) was used to localize Ig's in brains of animals inoculated with a cell associated strain of SSPE. Ig's were found in plasma cells in various stages of antibody production both in perivascular inflammatory lesions and scattered throughout the cerebral cortex. These findings offer corroborative evidence that the Ig found in SSPE ferret brain and CSF is actively synthesized within the CNS. Antibody was also demonstrated in glial and neuronal cell bodies and processes and in postsynaptic profiles. These are the same sites where measles virus antigens are most frequently found and suggests the possibility of immune complex formation.

Immunohistochemical findings in cranial arteritis.

The cause of cranial arteritis is unknown, but the demonstration of immunoglobulin and complement in temporal artery biopsies by immunofluorescence suggest that it may be a disease of disordered immunity. Because of the inevitable problems of histologic interpretation associated with the fluorescent technique, 15 temporal artery biopsies from patients with active arteritis were examined by an immunoperoxidase method. Varying amounts of IgA, IgG, and IgM were identified in plasma cells and macrophages. Extracellular IgG was identified in 1 case, but there was no staining for complement. These findings provide no support for the concept of cranial arteritis as a form of immune complex vasculitis.

Radioimmunometric demonstration of immunoglobulin G on cancer cells derived from malignant pleural effusions.

Well-preserved viable cancer cells, lymphocytes and mesothelial cells were prepared from various human malignant and non-malignant pleural effusions by a Ficoll-Hypaque discontinuous gradient. Using 125I-labelled protein A from Staphylococcus aureus as tracer, IgG was found on all cancer cells and also on apparently non-malignant mesothelial cells. Loss of cell surface IgG by overnight incubation, and fluctuation of surface IgG levels in the same patient occurred with cancer cells as well as with mesothelial cells. The IgG in vivo coat of cancer cells reflected the IgG levels of the sera rather than those of the pleural effusions. Binding of IgG from autologous and allogeneic sera and from supernatants of acid and mild overnight elution of cancer cells and mesothelial cells did not indicate a cancer-specific uptake of IgG. Various unspecific modes of IgG uptake by cancer cells are discussed including the possibility of a non-immune mechanism. Modification and loss of antigens by metastatic cells have also been considered.

Test systems for detection of human T associated Ig-related surface determinants using chicken antiglobulin reagents

In earlier experiments the mixed antiglobulin rosetting reaction (MARR) and the direct antiglobulin rosetting reaction (DARR) were found to be of similar sensitivity in demonstration of immunoglobulin (Ig)-bearing B lymphocytes. These test systems were used in the present study together with chicken anti-F(ab′) 2, anti-IgM and anti-IgA antibodies to detect Ig-related determinants on human peripheral blood T cells. Although for each antiglobulin reagent the reactivity of viable T cells in the MARR and the DARR appeared to be optimal after treatment of the lymphocytes with neuraminidase, the MARR was always more sensitive than the DARR. Further, when test conditions were established so as to avoid erroneous detection of Ig bound to T cell Fc receptors, the MARR was found to be more sensitive than indirect immunofluorescence. Under the most sensitive mixed antiglobulin rosetting conditions, the mean number of T cells expressing F(ab′) 2-, α- and μ-related determinants was 99%, 50% and 39% respectively. The complete inhibition of the MARR by human IgM when anti-IgM antibodies are used and the more than 70% inhibition by IgA when anti-IgA antibodies are used, largely exclude false positive rosette formation due to contaminating specificities directed against non-Ig determinants on T cells. Trypsin treatment of T cells removed all determinants recognized by anti-IgM and anti-IgA antibodies and these cells reexpressed these determinants during in vitro culture; this indicates that these determinants are T cell products and do not represent adsorbed Ig molecules.

Demonstration of immunoglobulin in the neighbourhood of advanced atherosclerotic plaques

A necropsy survey of inflammatory cellular infiltration associated with atherosclerosis showed that the degree of inflammation correlated with the severity of the atherosclerotic plaque and was greater in the presence of medial disruption. Sections were stained for IgG and IgM using the immunoperoxidase technique. Advanced plaques with medial disruption showed staining of the cytoplasm of plasma cells in the adventitia with IgG and IgM antisera. IgG was also demonstrated in the atheromatous material. The immunoglobulin production may have been stimulated by antigens formed in the advanced lesion. Such an allergic reaction might explain some of the complications of advanced atherosclerosis.

Familial Lichen Planus

To the Editor.— The cause of lichen planus remains obscure. Although many theories have been put forward to explain its cause, none has been widely accepted. A viral cause has long been suspected, and psychogenic influences seem to be of striking importance in many patients. The recent demonstration of immunoglobulins in lichen planus lesions 1 suggests an autoimmune pathogenesis for this disease. At present, however, there is little evidence to suggest that the disease is genetically determined. We recently had the opportunity of observing three patients who had lichen planus, each with a positive family history. Report of Cases.—Case 1.— A 16-year-old girl had multiple whitish streaks of six months' duration on the buccal mucosa. There was no involvement of the skin or nails. A biopsy specimen taken from the oral mucous membrane confirmed the diagnosis of lichen planus. While this patient was undergoing treatment, violaceous, flat-topped papular

Demonstration of immunoglobulin G in normal human epidermis by peroxidase-labeled antibody.

Cytoplasmic immunoglobulin G (IgG) in normal human epidermis was defined by a peroxidase-labeled antibody method. A correlation between cytoplasmic staining and the serum level of IgG was found. Epidermal cells containing IgG were not present when the serum level of IgG was less than 1000 microgram/ml.