Analysis of polymorphism in CD86, CTLA4, TNF and IL10 genes in Serbian patients with pemphigus

Uvod: Oboljenja iz grupe pemfigusa - pemfigus vulgaris (PV) i pemfigus foliaceus (PF) su autiomunska bulozna oboljenja kože i sluznica. U patogenezi su od značaja autoantitela na dezmogleine i druge molekule koja dovode do akantolize i formiranja intraepidermalnog rascepa. Za aktivaciju autoreaktivnih T-ćelija u pemfigusu neophodno je prepoznavanja dezmogleinskih peptida i angažovanje CD28 molekula, koji je najznačajniji kostimulatorni receptor u aktivaciji naivnih T ćelija. CD28 molekul se vezuje za B7 familiju molekula kao što su CD80 i CD86. Pored toga, B7 molekuli mogu da aktiviraju inhibitorni receptor na T-ćelijama, CTLA-4, koji kontroliše mehanizam održavanja tolerancije na sopstvene peptide. Na značaj genetskih faktora u patogenezi pemfigusa ukazuju prethodna istraživanja kojima su definisani određeni genetski faktori kao što su i polimorfizmi pojedinačnih nukleotida u genima molekula od značaja za patogenezu pemfigusa. Ciljevi istraživanja: Odrediti distribuciju alela u genima za molekule CD86, CTLA-4, TNF i IL-10 kod pacijenata sa PV i PF, kao i kod zdravih osoba (dobrovoljni davaoci krvi) u Srbiji, ispitati da li je neki od ispitivanih polimorfizama faktor rizika za nastanak oboljenja (PV i PF) i uporediti ih sa podacima iz literature koji su dobijeni ispitivanjem istih polimorfizama kod geografski i etnički različitih populacija. Pacijenti i metode: Istraživanje je obuhvatilo 61 bolesnika sa pemfigusom od kojih je PV imalo 48, a PF 13 bolesnika i 486 zdravih osoba-kontrola (dobrovoljni davaoci krvi). Dijagnoza pacijenata sa pemfigusom postavljena je na osnovu kliničke slike i testa direktne imunofluorescencije (DIF test). Za detekciju i analizu polimorfizama gena za CD86, CTLA-4, TNF i IL-10 korišćene su komercijalne optimizovane smeše specifičnih oligonukleotida i TaqMan proba obeleženih FAM i VIC fluorohromima i to: CD86 rs1129055, CTLA4 rs733618 i rs5742909, TNF rs1800629 i rs361525 i IL-10 rs1800896 i rs1800871 (PE, Applied Biosystems). Amplifikacija fragmenata DNK Real-time PCR metodom vršena je pomoću navedenih smeša komercijalnih oligonukleotida i Maxima™ Probe qPCR 2X Master Mix, (Fermentas) uzimajući u obzir preporuke proizvođača reagenasa...Introduction: Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are rare autoimmune blistering diseases with presumed T cell dependent pathology. Activation of naïve T cells is dependent on antigen recognition, subsequent signaling through the T cell receptor complex (signal 1), and various other interactions of T cells with antigen presenting cells that may be collectively designated as signal 2, which is unconditionally required for T cell activation both in response to infection and to autoantigens. Among the best described interactions contributing to signal 2 are those mediated by B7 family molecules such as CD80 and CD86 with their ligands, CD28, providing activation signals, and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), conferring inhibition. Single nucleotide polymorphisms (SNPs) within genes encoding those molecules may alter the signaling process. It is not known whether functional genetic polymorphisms within genes encoding the aforementioned proteins may increase risk for developing PV and PF and, if so, whether they might serve as biomarkers for susceptibility to these diseases. Aims of the investigation: To examine functional single nucleotide polymorphisms within CD86 (rs1129055), CTLA4 (rs733618 and rs5742909), TNF (rs1800629 andrs361525), and IL-10 (rs1800896 and rs1800871) genes in 61 pemphigus patients and 486 healthy controls, and to compare results in Serbian pemphigus patients and healthy controls with results obtained in different populations groups. Patients and methods: Our study included 61 pemphigus patients, among them 48 with PV and 13 with PF, treated at the Dermatovenereology Clinics of the Clinical Centre of Serbia. Patients were stratified according to clinical presentation of the disease as a PV or PF patients. Diagnosis in all patients was established by merging clinical findings, histopathologic analysis, and the results of direct immunofluorescence tests. Blood samples from 486 healthy blood donors were obtained from the National Blood Transfusion Institute..

Ecthyma gangrenosum in a patient with non-Hodgkin lymphoma

Introduction. Ecthyma gangrenosum is a rare disease of the skin that causes the localized necrosis of the skin and subcutaneous fat tissue, leading to the multiple ulcerations surrounded by local hyperaemia. The ulcerations are usually localized in groins, and perianal area. In the majority of cases ecthyma is caused by a Pseudomonas aeruginosa sepsis. The disease usually appears in immunocompromized, most frequently hematological patients. Case report. We presented a 78-year-old woman who had been treated for non-Hodgkin lymphoma for the last 18 years. She had recently been given cytotoxics which led to neutropenia. The patient suddenly developed high fever, chill and diarrhea, followed by ecthyma gangrenosum cutaneous lesions in groins, axillas, right side of the neck and umbilicus. Pseudomonas aeruginosa and Proteus mirabilis, that were sensitive to several antibiotics were isolated. The treatment included rehydratation, antibiotics, surgical debridement and regular dressing with antiseptics. The healing of all lesions was achieved after sixteen weeks of the treatment. Conclusion. If haemorrhagic- necrotic lesions of the skin are developed in immunocompromised, usually haematologic patients, an Ecthyma gangrenosum has to be considered immediately, material for identification of a cause has to be taken, followed by immediate administration of antibiotics effective against Pseudomonas aeruginosa. Surgical debridement and other therapeutic modalities are to be considered in some patients.

Atypical pyoderma gangrenosum in a patient with osteomyelofibrosis

Background. Atypical forms of pyoderma gangrenosum generally appear on the upper extremities; most frequently they are associated with myeloproliferative disorders, including osteomyelofibrosis. A response to systemic steroids is more pronounced than in classical form. Sometimes it may be the first sign of an underlying malignancy. Case report. We reported a patient with atypical pyoderma gangrenosum developed during the course of a myeloid malignancy - osteomyelofibrosis. The lesions occurred after a minor trauma. Painful blistering plaques, with an elevated, bluish-gray border were located on the dorsal aspect of hands. No skin malignancy was found. The lesions resolved rapidly to systemic steroids. Conclusion. Considering the unusual clinical presentation which makes the diagnosis difficult, as well as the fact that atypical forms of pyoderma gangrenosum can be the first sign of malignancies, especially myeloproliferative ones, recognizing this entity enables timely guiding future investigations toward their prompt detection

Analysis of polymorphism in CD86, CTLA4, TNF and IL10 genes in Serbian patients with pemphigus

Uvod: Oboljenja iz grupe pemfigusa - pemfigus vulgaris (PV) i pemfigus foliaceus (PF) su autiomunska bulozna oboljenja kože i sluznica. U patogenezi su od značaja autoantitela na dezmogleine i druge molekule koja dovode do akantolize i formiranja intraepidermalnog rascepa. Za aktivaciju autoreaktivnih T-ćelija u pemfigusu neophodno je prepoznavanja dezmogleinskih peptida i angažovanje CD28 molekula, koji je najznačajniji kostimulatorni receptor u aktivaciji naivnih T ćelija. CD28 molekul se vezuje za B7 familiju molekula kao što su CD80 i CD86. Pored toga, B7 molekuli mogu da aktiviraju inhibitorni receptor na T-ćelijama, CTLA-4, koji kontroliše mehanizam održavanja tolerancije na sopstvene peptide. Na značaj genetskih faktora u patogenezi pemfigusa ukazuju prethodna istraživanja kojima su definisani određeni genetski faktori kao što su i polimorfizmi pojedinačnih nukleotida u genima molekula od značaja za patogenezu pemfigusa. Ciljevi istraživanja: Odrediti distribuciju alela u genima za molekule CD86, CTLA-4, TNF i IL-10 kod pacijenata sa PV i PF, kao i kod zdravih osoba (dobrovoljni davaoci krvi) u Srbiji, ispitati da li je neki od ispitivanih polimorfizama faktor rizika za nastanak oboljenja (PV i PF) i uporediti ih sa podacima iz literature koji su dobijeni ispitivanjem istih polimorfizama kod geografski i etnički različitih populacija. Pacijenti i metode: Istraživanje je obuhvatilo 61 bolesnika sa pemfigusom od kojih je PV imalo 48, a PF 13 bolesnika i 486 zdravih osoba-kontrola (dobrovoljni davaoci krvi). Dijagnoza pacijenata sa pemfigusom postavljena je na osnovu kliničke slike i testa direktne imunofluorescencije (DIF test). Za detekciju i analizu polimorfizama gena za CD86, CTLA-4, TNF i IL-10 korišćene su komercijalne optimizovane smeše specifičnih oligonukleotida i TaqMan proba obeleženih FAM i VIC fluorohromima i to: CD86 rs1129055, CTLA4 rs733618 i rs5742909, TNF rs1800629 i rs361525 i IL-10 rs1800896 i rs1800871 (PE, Applied Biosystems). Amplifikacija fragmenata DNK Real-time PCR metodom vršena je pomoću navedenih smeša komercijalnih oligonukleotida i Maxima™ Probe qPCR 2X Master Mix, (Fermentas) uzimajući u obzir preporuke proizvođača reagenasa...Introduction: Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are rare autoimmune blistering diseases with presumed T cell dependent pathology. Activation of naïve T cells is dependent on antigen recognition, subsequent signaling through the T cell receptor complex (signal 1), and various other interactions of T cells with antigen presenting cells that may be collectively designated as signal 2, which is unconditionally required for T cell activation both in response to infection and to autoantigens. Among the best described interactions contributing to signal 2 are those mediated by B7 family molecules such as CD80 and CD86 with their ligands, CD28, providing activation signals, and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), conferring inhibition. Single nucleotide polymorphisms (SNPs) within genes encoding those molecules may alter the signaling process. It is not known whether functional genetic polymorphisms within genes encoding the aforementioned proteins may increase risk for developing PV and PF and, if so, whether they might serve as biomarkers for susceptibility to these diseases. Aims of the investigation: To examine functional single nucleotide polymorphisms within CD86 (rs1129055), CTLA4 (rs733618 and rs5742909), TNF (rs1800629 andrs361525), and IL-10 (rs1800896 and rs1800871) genes in 61 pemphigus patients and 486 healthy controls, and to compare results in Serbian pemphigus patients and healthy controls with results obtained in different populations groups. Patients and methods: Our study included 61 pemphigus patients, among them 48 with PV and 13 with PF, treated at the Dermatovenereology Clinics of the Clinical Centre of Serbia. Patients were stratified according to clinical presentation of the disease as a PV or PF patients. Diagnosis in all patients was established by merging clinical findings, histopathologic analysis, and the results of direct immunofluorescence tests. Blood samples from 486 healthy blood donors were obtained from the National Blood Transfusion Institute..

Primary systemic amyloidosis

Background. Systemic amyloidosis is a rare disorder which usually occurs in aged persons and has a poor prognosis. Systemic amyloidosis can be primary, occasionally associated with multiple myeloma, or secondary, associated with another disease. Case report. We presented a 72-year-old male patient with periocular purpura ("racoon sign") and waxy papules, petechiae and ecchymoses on the neck and thoracic area. Purpuric macules were present also on the lips and tongue which was edematous (macroglossia). The skin lesions occurred two years earlier, the patient lost more than 15 kilograms of the body mass for less than a year. Immunoelectrophoresis of urine and serum demonstrated the presence of immunoglobulin light chains of the circulating monoclonal protein. Histopathological examination of skin lesions showed Congo red positive deposits in the derm. Cardiac evaluation revealed the signs of heart failure, and renal evaluation revealed nephrotic syndrome, with excessive protein lost. He was treated with oral melphalan and prednisolone, and died 7 days after starting the therapy due to heart failure. Conclusion. This patient considered as a rare case with systemic amyloidosis highlights the importance of histopathological and physical examination in any cases with periocular purpura, petechiae, ecchymoses and macroglossia