Direct Immunofluorescence Technique Research Articles

Patterns of Periodic Acid–Schiff Staining in Subepidermal Bullous Disorders with Emphasis on Bullous Pemphigoid

Abstract Background: Direct immunofluorescence (DIF) including salt split skin technique is quintessential in the diagnosis of subepidermal bullous disorders. These techniques are expensive and require technical expertise, limiting their diagnostic utility. The property of periodic-acid–Schiff to stain the basement membrane and the patterns produced by it was observed in a cohort of subepidermal bullous disorders, to see if it offers an alternative cost-effective testing method. Materials and Methods: This was a retrospective, observational study conducted in the department of general pathology, in a tertiary care medical center, over a period of 4 years. Only those cases for which paraffin sections, DIF and direct salt split skin immunofluorescence technique were available were included. The levels of anti-bullous pemphigoid (BP) antibodies were collected where available. Results: Of the 21 cases analyzed, 15 were BP, two each of epidermolysis bullosa acquisita (EBA) and lichen planus pemphigoides (LPP), one each of porphyria cutanea tarda (PCT), and bullous systemic lupus erythematosus (SLE). All the cases of BP, LPP and EBA showed a floor pattern of staining with periodic acid–Schiff (PAS) stain. Roof pattern of staining was observed in bullous SLE and PCT. PAS was found to be useful in diagnosing BP, especially in conjunction with an eosinophil rich bulla and anti-BP antibodies. Conclusion: We found that PAS stain could be an adjunct to hematoxylin and eosin stain when the diagnosis of BP is in doubt. However, larger sample size is needed to study its utility in other subepidermal bullous disorders.

Immunohistochemical Evaluation of C4d and C3d Markers in Bullous Pemphigoid as a Substitute for Direct Immunofluorescence Technique

Immunohistochemical Evaluation of C4d and C3d Markers in Bullous Pemphigoid as a Substitute for Direct Immunofluorescence Technique

The lupus band test: A review of the sensitivity and specificity in the diagnosis of lupus erythematosus.

The lupus band test (LBT) is a direct immunofluorescence (DIF) technique which shows a band of localised immunoglobulins at the dermo-epidermal junction in lesional, non-sun-exposed skin of patients with both systemic and cutaneous lupus erythematosus (LE), and in perilesional skin of patients with systemic LE. However, low sensitivity and poor concordance between histological and clinical diagnoses warrant a review of the application of the LBT in the diagnosis of LE. To assess the sensitivity and specificity of the LBT in diagnosing LE following clinico-pathological correlation (CPC). All cases sent to our pathology department between 2011 and 2018 for DIF with a clinical query of LE were reviewed. Data collection included demographic details, pathology requests, histology and DIF reports, clinical reports and diagnoses, and serology. Of 256 histology requests, 9% (n=23) had a positive LBT. This was discordant with the prevalence of LE diagnosis, as 46.3% were diagnosed with LE following CPC. The sensitivity and specificity of the LBT for LE was 17.6% and 98.8% respectively, with a positive predictive value of 92.9% and negative predictive value of 58.2%. The LBT is not a sensitive diagnostic test for LE, but is highly specific, and should be considered as a supportive diagnostic tool for LE. This is the largest reported case series evaluating the efficacy of the LBT in the diagnosis of LE.

DP09 The lupus band test: a review of the sensitivity and specificity in the diagnosis of lupus erythematosus

Abstract The lupus band test (LBT) is a direct immunofluorescence (DIF) technique which shows a band of localized immunoglobulins at the dermoepidermal junction in lesional, non-sun-exposed skin of patients with both systemic (SLE) and cutaneous lupus erythematosus and in the perilesional skin of patients with SLE. However, low sensitivity and poor concordance between histological and clinical diagnoses warrant a review of the application of the LBT in the diagnosis of lupus erythematosus (LE). The aim of this study was to assess the sensitivity and specificity of the LBT in diagnosing LE following clinicopathological correlation (CPC). All cases sent to our pathology department between 2011 and 2018 for DIF with a clinical query of LE were reviewed. Data collection included demographic details, pathology requests, histology and DIF reports, clinical reports and diagnoses and serology. Of 256 histology requests, 9.0% (n = 23) had a positive LBT. This was discordant with the prevalence of LE diagnosis, as 46.3% were diagnosed with LE following CPC. The sensitivity and specificity of the LBT for LE were 17.6% and 98.8%, respectively, with a positive predictive value of 92.9% and negative predictive value of 58.2%. The LBT is not a sensitive diagnostic test for LE, but is highly specific and should be considered as a supportive diagnostic tool for LE. This is the largest reported case series to evaluate the efficacy of the LBT in the diagnosis of LE.

Kidney biopsy in subsaharan Africa

We carried out a retrospective and descriptive study on biopsies examined between January 2015 and December 2019, in the pathological departments of University Teaching Hospital of Bouaké and Cocody-Abidjan. The KB came from four countries (Côte d'Ivoire, Togo, Guinea-Conakry and Burkina Faso). Optical microscopy and/or direct immunofluorescence techniques were used. All biopsy samples including epidemiological, clinical and pathological data and an optical microscopy and/or direct immunofluorescence study were included. The parameters studied were indications for KB, epidemiological profile, clinic, proteinuria and pathological aspects. Over the study period, we collected 179 KB, i.e. 35.8 KB/year. The mean age of the patients was 32.9 ±13.8 years (range 11-70 years). The sex ratio (M/F) was 1.03. Pure nephrotic syndrome was the main indication (64.2%, n = 115) for KB, followed by impure nephrotic syndrome (11.7%, n = 21), acute renal failure (ARF) (7.8%, n = 14) and rapidly progressive glomerulonephritis (RPGN) (7.8%, n = 14). Glomerulonephritis (GN) occurred in 86% (n = 158), vascular nephropathy in 11.7% (n = 21) and tubulointerstitial nephritis in 2.2% (n = 4). The nephropathies were preferentially focal segmental glomerulosclerosis (34.6%, n = 62), nephroangiosclerosis (10.6%, n = 19), membranous GN (10%, n = 18), post-infectious GN (8.9%, n = 16) and lupus GN (7.3%, n = 13). The KB is an essential step in the diagnosis of nephropathies. Focal segmental glomerulosclerosis is frequent in our study. The establishment of a Kidney registry would allow better knowledge of renal pathologies in sub-Saharan Africa.

Direct Immunofluorescence in Cutaneous and Systemic Lupus Erythematosus: A Literature Review

Cutaneous manifestations of lupus erythematosus (LE) have a variety of clinical phenotypes and require proper investigation for diagnosis. Knowledge of the direct immunofluorescence (DIF) technique has improved and played an important role in the diagnosis of cutaneous LE. This review explores and summarizes reported DIF findings of each cutaneous LE variant. Historically, DIF findings of cutaneous LE have revealed deposits of multiple immunoreactants at the dermo-epidermal junction with either linear or granular patterns. Immunoglobulin M is the most common immunoreactant and DIF findings of cutaneous LE variants overlap. Therefore, diagnosis of cutaneous LE requires a combination of monitoring patient history, physical examinations and laboratory studies. This review helps interpret and better understand the application of DIF studies in cutaneous LE.

Extensive Phenotypic Characterization of T Cells Infiltrating Liver Metastasis from Colorectal Cancer: A Potential Role in Precision Medicine.

Colorectal cancer (CRC) is one of the most common cancers worldwide, with liver metastasis being its main cause of death. This study harvested fresh biological material from non-tumor and tumor tissue from 47 patients with CRC liver metastasis after surgery, followed by mechanical cellular extraction and stain-lyse-wash direct immunofluorescence technique. Here, 60 different T-cell populations were characterized by flow cytometry. Tumor samples were also subdivided according to their growth pattern into desmoplastic and non-desmoplastic. When we compared tumor versus non-tumor samples, we observed a significantly lower percentage of T-lymphocyte infiltration in the tumor in which the CD4+ T-cell density increased compared to the CD8+ T cells. T regulatory cells also increased within the tumor, even with an activated phenotype (HLA-DR+). A higher percentage of IL-17-producing cells was present in tumor samples and correlated with the metastasis size. In contrast, we also observed a significant increase in CD8+ follicular-like T cells (CD185+), suggesting a cytotoxic response to cancer cells. Additionally, most infiltrated T cells exhibit an intermediate activation phenotype (CD25+). In conclusion, our results revealed potential new targets and prognostic biomarkers that could take part in an algorithm for personalized medicine approaches improving CRC patients' outcomes.

Laboratory techniques for diagnosing Giardia parasite in dogs in Mosul City, Iraq

The results of the current study showed that the different techniques of microscopic examination recorded the highest rate of infection and diagnosis of Giardia in dogs with significant differences at the level of probability p<0.05 It reached 73.4%, followed by the rapid test technique and indirect enzyme linked immunosorbent assay (ELISA) with a rate of 14.2% and 12.2% respectively. With regard to determining the sensitivity and specificity of the rapid test and microscopic examination, a sensitivity of 19% and a specificity of 100% were recorded. The results, using the indirect ELISA showed an infection rate of 12.2% of the total samples. The results showed that the number of positive samples with this technique amounted to 12 samples positive for Giardia lamblia out of the total samples positive for microscopic examination. To confirm the diagnosis of giardiasis, the results using the direct immunofluorescence technique using acridine orange fluorochrome showed the appearance of a glowing brilliance, as the cysts appeared oval green glowing and the trophozoites appeared glowing green pear.

Risk of Developing Asthma After Lower Respiratory Tract Infections with Respiratory Syncytial Virus During Childhood

Background: Recent studies indicate causal relationship between infection by respiratory syncytial virus (RSV) and bronchial asthma. We evaluated the incidence of bronchial asthma in children with RSV positive infection early in their childhood in a nation-wide cohort study. Methods: Children (aged between one month and 15 years) were evaluated for the presence of RSV infection when they presented with one or more acute respiratory tract infection symptoms (fever, cough, cold and wheezing) in a major tertiary care hospital in the Kingdom of Bahrain during a period of seven years. RSV detection was done using nasopharyngeal secretion (NPS) samples by direct antigen detection immunofluorescence technique. Number of children who were later diagnosed with asthma was recorded. Serum IgE levels were estimated. Risk of developing bronchial asthma is represented using relative risk (RR) [95% CI]. Children with asthma without prior RSV infection from the same population formed the historical control. Results: A total of 3782 children diagnosed with respiratory tract infection were recruited. We observed that RSV infection at younger age (during infancy) and severe infection were significantly associated with asthmatic episodes RR [95% CI]: 7 [5.5, 8.2]. Additionally, asthmatics with prior RSV infection had significantly higher total IgE levels (167 ± 37 IU/ml) compared to those without RSV infection (92 ±17 IU/ml). Mean (SD) age of children developing asthma with prior RSV infection was 0.7 (0.42) years compared to the historical control [6.8 (3.8) years] and was statistically significant. Conclusion: Infants with RSV infection have an increased risk of developing bronchial asthma later in the childhood. The more severe the RSV infection, the greater is the severity of bronchial asthma as indicted by serum IgE levels. Asthma in children with RSV infection occurs at much younger age compared to those without RSV infection.

Clinicopathological Features of Lupus Nephritis Patients in North-East India; A Single Center Retrospective Observational Study

Clinicopathological presentation of lupus nephritis (LN) patients varies with different race and ethinicity of the population. Only few studies describe clinicopathological spectrum of LN patients in the Indian population. The aim of this study was to determine the clinicopathological spectrum of LN in the North-East Indian population. This was a retrospective observational study that included patients with LN at a tertiary care center in North-East India from March 2007 to August 2018. Clinical and histopathological data at the time of presentation were collected from hospital records. Renal biopsies were examined by light microscopy and direct immunofluorescence techniques. A total of 340 patients of LN were included in this study. The mean age of presentation was 22.42 ± 4.3 years. The minimum age at presentation was 8 years and 18.8% belonged to the <18 year age group. The present study showed a male:female ratio of 1:8. The majority of patients were of class IV (71.9%). Arthralgia (47.1%) and anemia (60.3%) were the most common presenting symptom and sign, respectively. Immunoglobulin (Ig) G was the most abundant immunoglobulin (positive in 98.47%) and the least positive was IgA (positive in 41.18%). Complement (C) 3 and C1q were positive in all. Full house deposition was found in 59.3% of the biopsies. The rates of hypertension, microscopic hematuria, renal dysfunction, and nephrotic syndrome were 43.5, 59.12, 45.9, and 35.3%, respectively. Patients of LN in the North-East Indian population present at an earlier age with a more severe form of the disease (class IV) at the time of presentation.

Study of the Relationship between Vitamin D Level and the Increase in the Severity of Covid-19 Infection in Kirkuk City

The study aimed to find the relation between Covid-19 infection and reduction in vitamin-D levels andwith development of severe infection The study was conducted in Kirkuk city during July 2020, included120 COVID-19 patients who diagnosed by collecting of naso/oropharyngeal swabs and the virus RNAwas qualitatively detected by real-Time PCR in Public Health Laboratory according manufactureinstruction and WHO protocol. The study also included 30 healthy persons as control group. The studyincluded the collection of blood samples for determination of vitamin-D level (I-chroma, Korea) by directimmunofluorescence technique and according to manufacture instruction. The study also included collectionof associated information like age, sex, residence, BMI and by contacting with all patients daily until the10th day of infection to know their status after treatment. The study showed that, majority of patients wereaged from 22 to 41 years of old (P. value <0.001). The study showed that 62.5% of COVID-19 patientswere males compared with 37.5% females (P<0.05). The study showed that were asymptomatic, 44.17% ofcases of COVID-19 patients were with mild infection and 10% were with severe infection (P<0.001). Thestudy showed a significant relation between Covid-19 infection status with age as the highest mean of ageof Covid-19 infected patients was recorded in those who with severe infection. The study showed that thelowest mean of serum Vitamin was found in COVID-19 patients comparing with healthy control (12.8±3.6v.s. 29.3±3.1 ng/ml) (P: <0.001). The study also demonstrated that the level of vit. D was significantlyreduced in severe infected COVID-19 patients followed by patients with moderate infection and the highestmean was in patients with asymptomatic infection (P<0.001).

Conceptualization and validation of an innovative direct immunofluorescence technique utilizing fluorescein conjugate against IgG + IgG4 for routinely diagnosing autoimmune bullous dermatoses.

IntroductionAutoimmune bullous diseases (ABDs) are potentially life-threatening mucocutaneous illnesses that require diagnosis with direct immunofluorescence (DIF). In this study we compared the diagnostic accuracy of traditional DIF (DIFt; separate immunoglobulin (Ig) G, IgG1, IgG4, IgA, IgM and C3 deposits detection) and modified DIF (DIFm; simultaneous IgG + IgG4 deposits detection instead of separate IgG and IgG4 deposits detection) in routine diagnostics of ABDs.Material and methodsEighteen patients with ABDs (7 with pemphigus dermatoses and 11 with subepithelial ABDs) were evaluated with DIFt and DIFm.ResultsThe agreement of detectability of IgG immunoreactants was obtained in 16 ABD cases (88.89%), as positive results in both DIFt and DIFm were obtained in 13 cases and negative results in both DIFt and DIFm were obtained in 3 cases. One ABD case (Brunsting-Perry pemphigoid) (5.56%) was negative in DIFm with a positive DIFt result (IgG1 deposits). One ABD case (bullous pemphigoid) (5.56%) had only C3 deposits in DIFt with a positive DIFm reading (IgG + IgG4 deposits). A statistically significant relationship (p = 0.0186) between DIFm and DIFt results was revealed using Fisher’s exact test.ConclusionsBoth DIFt and DIFm are useful methods to detect deposition of IgG immunoreactants, but it seems that the innovative DIFm method slightly increases the detectability of IgG/IgG4 immunoreactants in relation to DIFt. The introduction of DIFm into routine laboratory diagnostics of ABDs seems to be justified, as it enables the abandonment of separate FITC conjugates for IgG and IgG4, which is important for cost-effectiveness.

Equine Rabies in the Southern Region of Piauí State

Background: Rabies is an infectious disease that is important in the "One Health" worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil.Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2oC, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4oC to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with antigen-labeled antibodies were performed in the imprint lamina of these fragments. The fragments were treated according to specific protocol. The results were negative for DIF in the collected equine fragments. For complementary exam, the samples were homogenized, clarified and inoculated intracranial in BALB/C mice, being observed for up to 30 days. The samples were positive after the bioassay.Discussion: Piauí is a state with great equestrian activity that expose the animals to the risks of transmission of infectious diseases. Among these diseases, rabies is important for affecting horses, but also humans (veterinarians and owners). In the present report, the equine showed clinical signs of furious rabies for a short period, rapidly evolving to paralytic form. This clinical aspect must be carefully evaluated by the veterinarian, in order to avoid false clinical suspicions such as tetanus or other non-infectious diseases. The official diagnosis of the rabies is DIF technique, with high sensitive (80-100%). According to the results of the DIF technique, it was possible to confirm the clinical suspicion of rabies in mice previously inoculated with emulsion of fragments of the equine central nervous system (CNS). This fact demonstrates that in negative results for CNS samples from the horse, the bioassay increases the sensitivity of the test and avoids false negative diagnoses. Thus, it was possible to prove that rabies is affecting the equines in the southern region of Piauí state and alerts the breeders and the community to intensify surveillance and control of the hematophagous bats. For the authors, this is the first scientific report of rabies in the region studied.

Uses of direct and indirect immuno-fluorescent techniques for demonstration of nematodes infection in sheep in Nineveh government

Uses of direct and indirect immuno-fluorescent techniques for demonstration of nematodes infection in sheep in Nineveh government

Rowell syndrome in dermatological practice (a clinical case)

Rowell syndrome is a rare cluster of symptoms characterized by clinical manifestation of lupus erythematosus and erythema multiforme (EM). About 100 cases of the syndrome have been reported in medical publications during the last 100 years. This may be related to misinterpretation of the symptoms and subsequent incorrect diagnosis due to its EM-like manifestations. Important clues for the diagnosis of Rowell syndrome are findings of positive rheumatoid factor, anti-nuclear antibodies and other erythematoid markers, as well as additional investigations, in particular, direct immunofluorescence technique. The paper describes a clinical case of Rowell syndrome in a 16-year old male patient. The diagnosis was challenging due to EM-like skin manifestations and required additional laboratory work-up, as well as the patient's follow-up. The diagnosis of Rowell syndrome was based on the clinical manifestations and on such diagnostic criteria as positive rheumatoid factor and anti-nuclear antibodies, as well as histological and laboratory abnormalities characteristic of the erythematosis. The patient was hospitalized and received the following treatment: prednisolone infusion (2.5 mg/kg/daily for 7 days), chloropyramine (1 mL i.m. twice daily for 5 days), hydroxychloroquine (6.5 mg/kg daily for 5 days), magnesium asparaginate/potassium asparaginate (one tablet (166.3 mg/175 mg) 3 times daily for 7 days), topical methylprednisolone aceponate cream 1% (once daily for 7 days). The treatment resulted in positive changes in the skin lesion and improvement of his general state. This clinical observation gives an example of classic Rowell syndrome proven both by lab and clinical signs, taking into account skin symptoms of lupus erythematosus and EM-like rash.

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.

EVALUATION OF BONE MARROW INFILTRATION BY MULTIDIMENSIONAL FLOW CYTOMETRY IN PATIENTS WITH DIFFUSE LARGE B‐CELL LYMPHOMA: SUB‐STUDY OF A PHASE 2 GELTAMO CLINICAL TRIAL

Introduction: Multidimensional flow cytometry (MFC) has not yet shown a high clinical value in patients with diffuse large B-cell lymphoma (DLBCL), probably due to the phenotypic heterogeneity of these lymphomas. In the present study, we have evaluated bone marrow (BM) infiltration in patients with DLBCL using MFC with the following objectives: 1) evaluating its prognostic impact compared to other techniques such as histology or PET; 2) determine the specific phenotypic pattern of the tumor cell of each patient for subsequent follow-up of minimal residual disease (MRD) after treatment. Methods: We centrally evaluated BM samples from patients included in the multicenter randomized phase 2 clinical trial GEL-R-COMP-2013 (EudraCT: 2013-001065-17), that compares R-CHOP versus R-COMP in patients with newly diagnosed DLBCL or follicular lymphoma (FL) grade 3b. High resolution (8 colors) and high sensitivity (more than 1 million cells analyzed) direct immunofluorescence techniques were used, following the protocols defined by EuroFlow. An automatic analysis was carried out in addition to the manual analysis, using the Compass III tool, available in the Infinicyt analysis software (Cytognos, S.L.). Results: From a total of 91 patients included in the clinical trial, 54 participated in the present study (48 DLBCL and 6 FL grade 3b). The median age was 75 years (61-86). BM infiltration was detected by MFC in 20 patients (37%) at diagnosis and 0 out of 13 at post-treatment evaluation. Regarding the phenotypic characterization, concordant BM infiltration by DLBCL was detected in only 6 cases. In contrast, discordant infiltration was detected in 14 (70%) patients, with a very heterogeneous phenotype: 3 low grade FL, 2 chronic lymphocytic leukemia, 5 marginal zone lymphoma and 4 small cell lymphoma with non-specific phenotype. Molecular studies showed monoclonal rearrangements in 14 out of 17 (82.3%) samples tested. Preliminary analysis of the prognostic impact of BM infiltration by MFC is shown in Table 1. The presence of concordant BM infiltration was associated with worse survival, although the differences were not statistically significant, possibly due to the low number of cases. On the other hand, discordant BM infiltration had no significant prognostic influence. Conclusions: High resolution MFC allowed the detection of small clonal populations in BM in a very high proportion of patients with newly diagnosed DLBCL. In most cases, infiltration was discordant with tumor histology and, apparently, had no prognostic relevance. MRD was not detected in any of the cases. We plan to correlate MFC findings with those of histology and PET, also centrally reviewed, as well as to perform molecular studies on lymph node and BM samples to assess their clonal relationship. Keywords: diffuse large B-cell lymphoma (DLBCL); flow cytometry. Disclosures: Sancho, J: Honoraria: Roche, Janssen, Gilead, Khen Fharma, Celgene, Sanofy, Servier, Mundipharma; Research Funding: Gilead. Martín, A: Consultant Advisory Role: Celgene, Roche.; Honoraria: Celgene, Roche, Servier, Janssen.

Complications of Urogenital Chlamydial Infection in Women

The objective of the research was to identify the spectrum of complications in women with chronic urogenital chlamydial infection.
 Materials and methods. There were examined 128 women with chronic inflammatory diseases of genital organs at the age of 16-40 years who were diagnosed with urogenital chlamydial infection and 25 apparently healthy women. In both women with chlamydial infection and healthy ones, urogenital chlamydial infection was diagnosed based on the data of clinical examination and the results of laboratory tests (the identification of chlamydial morphological structure on the pathologic specimens stained according to the Romanowsky-Giemsa method; the identification of chlamydial antigens using the direct immunofluorescence technique; the study of Chlamydia trachomatis antibody titers using the enzyme-linked immunosorbent assay).
 Results. According to the results of our study, chlamydiae were the most common causes of inflammatory lesions of the urogenital organs in women of different ages leading to different reproductive complications and affecting females mostly at the age of 21-30 years. In women of Group I and Group II, chronic chlamydial disease was detected; disease duration ranged from 6 months to more than 2 years. Miscarriages, infertility (primary, secondary), ectopic pregnancy (tubal, ovarian) were the severest and the most numerous reproductive complications in the examined patients.
 Conclusion. Chronic chlamydial infection is the most common disease of the female urinogenital organs leading to a wide spectrum of complications including infertility (primary, secondary), miscarriages, ectopic pregnancy (tubal, ovarian), chronic abdominal pain, sexual dysfunction (low libido, hypo/anorgasmia, painful intercourse, neurotic symptoms).

Oral Lichen Planus - Known and Unknown: a Review.

Lichen planus is a chronic mucocutaneous inflammatory disease aff ecting 1-2% of the general population with maximum prevalence of the disease in women above the age of 40. Its aetiology remains unclear and the pathogenesis is still the object of much speculation. It is considered to be an autoimmune disorder mediated mainly by the T-lymphocytes. The present paper presents the most well-known external agents (viruses in particular), internal agents like stress, and the heat shock protein thought to be trigger factors and describes the action of diff erent cells and proteins associated with the development of that disease. Diagnosis is based on clinical and histopathologic evidence; direct and indirect immunofluorescence techniques can also be of use. Despite the wide variety of therapeutic modalities, treatment outcomes are often insufficient. Currently, topical corticosteroids are widely accepted as a standard therapy, but also retinoids, calcineurin inhibitors and other immunosuppressants can be administered. Because of the aspect relevant to these drugs, priority is given to alternative harmless methods such as LLLT and PDLT. There is an ongoing controversy in the literature about the possible premalignant character of oral lichen planus, however, periodic followup is recommended.

Toll-Like Receptors (TLRs) Play Role in Adaptive Immunity in Rabbits Immunized by Sarcoptes scabiei Proteins

Sarcoptic mange is one of the most economically important diseases in goats in Indonesia, and increasing number of cases of treatment failure is being reported because of drug resistance. Nowadays, it is considered as an emerging/re-emerging parasitic disease that threatens human and animal health globally. Toll-Like Receptors (TLR) is a receptor which plays role in innate immunity due to microbial infection. TLR-5 is a receptor that can recognize ligand produced by bacterial component, and TLR-7 is involved in the recognition of ligand that similar to ssRNA virus. This research aims are to detect Sarcoptes scabiei antigenic protein which can induce cellular immune response in rabbit as adaptive immunity with TLR-5 and TLR-7 as marker.This research was performed in several stages: isolation and identification S. scabiei from scabies infected goats; extraction of soluble protein S.scabiei mite, the rabbit immunization by inoculating protein antigen S. scabiei with dosage of 500µg, repeated five times as booster in two weeks, the examination of TLR-5 and TLR-7 expression using direct immunofluorescence technique. The result of cellular immune response is shown by TLR-5 and TLR-7 expression in rabbit T limphocytes which appear yellow to green fluorescence color using fluorescence microscope. The amount of fluoresnce T lymphocytes showed a significant difference (p<0.05) between control and various boosters, and significantly increased in 3rd booster or 42 days post immunization. The conclusion showed protein from S. scabiei mites taken from goat contains ligands, acting as receptors which involve in pathogen associated molecular pattern (PAMP) that can induce adaptive immunity and recognized by TLR-5 and TLR-7. It shows that TLR is not only involved in innate immunity but also in adaptive immunity, and can be used as alternative adjuvant development. Keywords: Sarcoptes scabiei, TLR- 5, TLR-7, goats, rabbits.