[Analysis of IFN-γR1 (CD119) and IL-12Rß1 (CD212) Deficiency by Flow Cytometry]. / IFN-γR1 (CD119) ve IL-12Rß1 (CD212) Eksikliginin Akan Hücre Ölçer ile Analizi.

Mendelian susceptibility to mycobacterial diseases (MSMD) is a rare primary immune deficiency (PID). IL-12Rß1 deficiency is the most frequently observed of more than 16 genetic defects that have been identified for MSMD. Genetic and immunological tests are remarkable in the diagnosis of PID. In this study, it was aimed to determine the expression of IFN-γR1 and IL-12Rß1 in patients with MSMD, their relatives, and healthy individuals and to evaluate the importance of flow cytometry as a fast and reliable method in the diagnosis of MSMD. IFN-γR1 and IL-12Rß1 expression levels were analyzed in 32 volunteers including six patients, six relatives, and 20 healthy individuals. The normal range of IFN-γR1 and IL-12Rß1 levels among healthy individuals were determined. IL-12Rß1 expression level in lymphocytes was found to be low in one patient's relative, and less than 1% in three patients and in one patient's relative. It was observed that the IL-12Rß1 expression levels of the patient with STAT1 deficiency were increased compared to the healthy individuals. No difference was found in the expression levels of IFN-γR1 and IL-12Rß1 in one patient, but IFN-γR1 expression was decreased in one patient compared to healthy individuals. Our results show that the determination of IL-12Rß1 and IFN-γR1 deficiencies by flow cytometry can be used as a rapid and reliable method for the diagnosis of MSMD. The use of this method as a screening test will enable early diagnosis especially in patients whose genetic diagnosis has not been confirmed and clinically compatible with MSMD. In addition, it is thought that IL-12Rß1 and IFN-γR1 range data obtained from healthy individuals will be considered as a reference source in routine and research studies to be conducted with MSMD.

Immune modulation as a consequence of SARS-CoV-2 infection.

Erroneous immune responses in COVID-19 could have detrimental effects, which makes investigation of immune network underlying COVID-19 pathogenesis a requisite. This study aimed to investigate COVID-19 related alterations within the frame of innate and adaptive immunity. Thirty-four patients clinically diagnosed with mild, moderate and severe COVID-19 disease were enrolled in this study. Decreased ILC1 and increased ILC2 subsets were detected in mild and moderate patients compared to healthy controls. NK cell subsets and cytotoxic capacity of NK cells were decreased in severe patients. Moreover, CD3+ T cells were reduced in severe patients and a negative correlation was found between CD3+ T cells and D-dimer levels. Likewise, moderate and severe patients showed diminished CD3+CD8+ T cells. Unlike T and NK cells, plasmablast and plasma cells were elevated in patients and IgG and IgA levels were particularly increased in severe patients. Severe patients also showed elevated serum levels of pro-inflammatory cytokines such as TNF-α, IL-6 and IL-8, reduced intracellular IFN-γ and increased intracellular IL-10 levels. Our findings emphasize that SARS-CoV-2 infection significantly alters immune responses and innate and acquired immunity are differentially modulated in line with the clinical severity of the disease. Elevation of IL-10 levels in NK cells and reduction of CD3+ and CD8+ T cells in severe patients might be considered as a protective response against the harmful effect of cytokine storm seen in COVID-19.

Genotypic characterization of Pseudomonas aeruginosa isolates from Turkish children with cystic fibrosis.

Objective: To identify epidemic and other transmissible Pseudomonas aeruginosa strains, genotypic analyses are required. The aim of this study was to assess the distribution of P. aeruginosa strains within the Turkish pediatric cystic fibrosis (CF) clinic population. Methods: Eighteen patients attending the pediatric CF clinic of Cerrahpasa Medical Faculty were investigated in the study. Throat swab and/or sputum samples were taken from each patient at 3-month intervals. The isolates of patients were analyzed by pulsed-field gel electrophoresis (PFGE). The intra- and interpatient genotypic heterogeneity of isolates was examined to determine the clonal isolates of P. aeruginosa within the cohort. Results: A total of 108 clinical isolates of P. aeruginosa were obtained from 18 patients between May 2013 and May 2014. The pulsotypes of the first patient's isolates could not be obtained by PFGE. From the remaining 17 patients and 101 isolates, 55 distinct pulsotypes were detected. The number of pulsotypes observed in more than one patient (minor clonal strains, cluster strains) was 8 (14.5%), and one of them colonized three patients. However, none of them was detected in more than three patients. These pulsotypes were composed of 20 isolates. In addition, with the PFGE analysis of 81 isolates, we detected 47 (85.6%) pulsotypes, which belonged to only one patient. Over different periods of this study, only 2 (11.8%) patients were colonized with the same pulsotype. Conclusion: Our study indicates that there was considerable genomic diversity among the P. aeruginosa isolates in our clinic. The presence of shared pulsotypes supports cross-transmission between patients.

Simple Identification of Mycobacterial Species by Sequence-Specific Multiple Polymerase Chain Reactions.

Several species of mycobacteria cause infections in humans. Species identification of clinical isolates of mycobacteria is very important for the decision of treatment and in choosing the appropriate treatment regimen. We have developed a multiplex PCR method that can identify practically all known species of mycobacteria, by determination of single-nucleotide differences at a total of 13 different polymorphic regions in the genes of rRNA and hsp65, in four PCR mixes. To achieve this goal, single-nucleotide differences in these polymorphic regions were used to divide mycobacterial species into two groups, than four, eight, etc., in an algorithmic manner. It was sufficient to reach single species level by evaluating 13 polymorphic regions. Evaluation of the multiplex PCR patterns by observable real-time electrophoresis (ORTE) simplified species identification. This new method may enable easy, rapid, and cost-effective identification of all species of mycobacteria.

Inflammatory bowel disease and mycobacteria: how much can we trust isoniazid prophylaxis during antitumor necrosis factor therapy?

OBJECTIVES: Isoniazid (INH) prophylaxis is recommended for the prevention of tuberculosis (TB) reactivation before or/and during initiation of treatment with tumour necrosis factor antagonists (anti-TNF agents). Nonetheless, the long-term effectiveness of chemoprophylaxis is not clear. In this study, we aimed to evaluate the characteristics of patients who developed TB reactivation in spite of INH prophylaxis associated with anti-TNF treatment. PATIENTS AND METHODS: In this retrospective study, medical records of 1263 patients with inflammatory bowel disease were reviewed. Baseline TB screening tests (purified protein derivative test and/or QuantiFERON-TB Gold test) were performed on all patients before initiation of anti-TNF therapy. Patients with purified protein derivative of more than 5 mm and/or a positive result of the QuantiFERON-TB Gold test received INH prophylaxis for 9 months. We analysed the data of patients diagnosed with TB reactivation during the anti-TNF treatment despite INH chemoprophylaxis. RESULTS: Overall, 175 patients underwent anti-TNF treatment. Sixty of these 175 patients had pretreatment testing showing latent TB infection and therefore were treated concomitantly with INH for 9 months in addition to their anti-TNF treatment. TB reactivation occurred in four of these 60 co-INH/anti-TNF treated patients. Active TB was diagnosed after 37.5±27 (range: 18-84) months of anti-TNF treatment. In two of the four patients that active TB was diagnosed, was also detected other Mycobacterium spp.: M. bovis in one patient and M. genavense in the other one. CONCLUSION: INH chemoprophylaxis may not prevent the reactivation of TB during anti-TNF therapy in the long-term. Patients should be carefully and periodically screened for TB reactivation during anti-TNF therapy.

Inherited IL-12Rß1 Deficiency in a Child With BCG Adenitis and Oral Candidiasis: A Case Report.

Tuberculosis is a major worldwide problem, and protection from it is achieved mainly by live attenuated bacille Calmette-Guérin vaccine, which is capable of causing disease in immunocompromised host. Oral thrush is abnormal in healthy children, which suggests an underlying immunodeficiency. Mendelian susceptibility to mycobacterial disease is a rare primary immunodeficiency characterized by a selective predisposition to weakly virulent Mycobacteria and Salmonella and also predisposition to chronic mucocutaneous candidiasis. Interleukin 12 receptor ß1 (IL-12Rß1) deficiency is the most common disease of Mendelian susceptibility to mycobacterial disease, and to date only 50 IL-12Rß1 deficient patients with clinical signs of chronic mucocutaneous candidiasis have been reported. We report a 2.5-year-old daughter of consanguineous parents with both regional bacille Calmette-Guérin lymphadenitis and recurrent oral candidiasis carrying biallelic R175W mutation in the IL12RB1 gene, resulting in complete loss of expression of IL-12Rß1. To our knowledge, this is the first report of bacille Calmette-Guérin lymphadenitis with concurrent oral candidiasis displaying such a mutation. New mutations and wide clinical diversities are the indisputable fact of populations with a high rate of consanguineous marriages.

Comparison of tuberculin skin test and QuantiFERON®-TB Gold In-Tube for the diagnosis of childhood tuberculosis.

BACKGROUND: Tuberculosis (TB) is an important worldwide ongoing health issue. To be able to control TB, one should not only cure active TB but also identify childhood TB patients who have the possibility to develop active disease in the future. The aim of this study was to compare a century-old tuberculin skin test (TST) and QuantiFERON-TB Gold In-Tube (QFT-GIT) test, developed as an alternative to TST and which has been claimed to be superior to TST in several ways, in the diagnosis of childhood TB. METHODS: Fifty-three children with TB between 5 months and 17.5 years of age and 92 healthy children from the same age group with no risk factors for TB were recruited into the study. All children underwent TST and QFT-GIT test and their demographic, clinic and laboratory data were recorded. Data were analyzed using SPSS 14. RESULTS: A total of 53 patients were diagnosed with TB. Mean patient age was 8.5 ± 4.3 years (range, 5 months-17.5 years). A total of 41.7% of the patients were female. Sixteen of 53 patients had confirmation on culture. QFT-GIT test was positive in 16 and TST was positive in 15 among 16 children with culture-confirmed TB. The sensitivity of TST and QFT-GIT were 93.8% and 100.0%, and the specificity of TST and QFT-GIT were 100.0% and 97.8%, respectively. With regard to the 53 TB children including those without bacteriological confirmation, QFT-GIT was positive in 33 children, and TST was positive in 44 children. The sensitivity of TST and QFT-GIT was then 83.0% and 62.3%, and the specificity, 100.0% and 97.8%, respectively. CONCLUSION: Although positive QFT-GIT test is very significant for TB, negative results will not exclude TB infection. TST and QFT-GIT used together may provide more efficient results.

Non-tuberculous mycobacteria infection: 75 cases.

INTRODUCTION: Non-tuberculosis mycobacterium is especially seen in AIDS and non-immunosuppressant patients. This study was designed to evaluate data relating to non-tuberculosis mycobacterium content in patients' sputum for the clinical importance. PATIENTS AND METHODS: During 2009-2010 at Sureyyapasa Chest Diseases and Chest Surgery Training and Research Hospital clinics, 75 patients [30 women (40%) and 45 men (69%); mean age (SD): 48.7 (15.9) years] with non-tuberculosis mycobacterium were determined by the rapid test and NAP test in Mycobacteria Growth Indicator Tube (MGIT), which had atypical growth in 51.864 Lowenstein-Jensen. Identification was done with Hsp65PCRREA methods in 32 (43%) cases. Treatment management, radiology, bacteriology, co-morbidity, treatment outcomes were evaluated from medical records, calling patients and from tuberculosis dispensaries. RESULTS: In 9 (28%) patients Mycobacterium abscessus, in 8 (25%) patients Mycobacterium avium complex (MAC), in 5 (16%) patients Mycobacterium kansasii was found with identification Hsp65PCRREA methods. In 18 (24%) of 75 cases with American Thoracic Society definition and treatment criteria, treatment was administered using major and minor drugs. Standard tuberculosis treatment was administered in 25 (33%) of the 75 cases. In 8 of 25 (32%) cases identification of non-tuberculosis mycobacterium was evident. In 32 of 75 cases follow up was performed with no treatment. One positive atypical growth culture was identified in 23 (72%) of 32 patients. Treatment was administered in 43 cases while 25 (58%) of 43 were cured, 3 (7%) of 43 were default and 3 (7%) died. Drug resistance was the outcome in 36 cases. While 31 (86%) had any drug resistance, 27 (75%) had HR drug resistance. Past history of tuberculosis treatment was evident in 20 (40%) cases. Respiratory and non-respiratory diseases were identified equally in 18 (38%) cases. Radiological consolidation in 28 (65%), and cavity in 16 (37%) cases were determined. CONCLUSION: In order to carry out the right treatment and epidemiologic evaluation, it is important to identify non-tuberculosis mycobacterium by culture methods.

Treatment with Mycobacterium vaccae ameliorates airway histopathology in a murine model of asthma.

The objective of this study was to evaluate the effect of intratracheal (i.t.) or subcutaneous (s.c.) Mycobacterium vaccae treatment on lung histopathology and cytokine responses in a murine model of asthma. BALB/c mice were divided into four groups. To establish an asthma model, Groups I, II and III received intraperitoneal (i.p.) ovalbumin (OVA) and were challenged with i.t. OVA three times (days 41-47). On the same days, mice in Groups I and II were treated with i.t. and s.c. Mycobacterium vaccae, respectively. Mice in Group IV served as controls. On day 49, lungs were taken out for histopathological evaluation. Cytokine levels were determined in splenocyte culture supernatants by ELISA. The thickness of basement membrane and hyperplasic goblet cells in small airways were found to be significantly more in Group III than Group I. Furthermore, smooth muscle and epithelial thickness in small and large airways and hyperplasic goblet cell numbers in all sized airways of this treatment group were not significantly different from controls. Epithelial thickness in medium and large airways, hyperplasic goblet cells in all sized airways, and basement membrane in small and large airways were not significantly different in Group II when compared to controls. OVA-stimulated IL-5 levels was significantly higher in Group I when compared to Group III. OVA-stimulated IL-5 and spontaneous IL-5 levels were significantly higher in Group II than Group III. We demonstrate that subcutaneous and intratracheal Mycobacterium vaccae administered along with allergen has an ameliorating effect in the modulation of airway histopathological changes in OVA sensitized mice.

Frequency of Chlamydia pneumoniae and Mycoplasma pneumoniae infections in children.

OBJECTIVE: Chlamydia pneumoniae and Mycoplasma pneumoniae are among the most important pathogens of acute respiratory infections in children between the ages of 5 and 15 years. We aimed to investigate seasonal frequency of respiratory infections caused by C. pneumoniae and M. pneumoniae, frequency of coinfection, clinical findings and to determine relationship between clinical findings and laboratory results. MATERIAL AND METHODS: Total of 284 patients (ranging 5-15 years of age), admitted to out-patient clinic with symptoms of respiratory tract infections between January 2004 and June 2005, were enrolled in the study. IgA, IgG and IgM antibodies against C. pneumoniae were quantitatively detected in all serum samples by using microimmunofluorescence (MIF). For the M. pneumoniae infection an IgM titer in the ELISA test were analyzed. Nasopharyngeal smear samples were collected for PCR detection. RESULTS: Mean age was 8 +/- 2.2 (range 5-14) years. Mycoplasma pneumoniae IgM in 86 (30.2%) cases, C. pneumoniae IgM in one (0.3%) case, IgA in six (2.1%) cases and IgG in 10 (3.5%) cases were found positive. In 10 (3.5%) cases, both C. pneumoniae IgG (a titer of >1/216) and M. pneumoniae IgM were found positive concomitantly. The M. pneumoniae IgM in winter was found significantly higher compared to other seasons. Mycoplasma pneumoniae PCR method was performed on a total of 203 samples in 33 (16.2%) of which M. pneumoniae was found positive. The false positive ratio of PCR technique was found 16.2%. In a total of 217 examined samples by PCR method, the DNA of C. pneumoniae was found positive in two patients. CONCLUSION: Mycoplasma pneumoniae was a common pathogen in respiratory infections. The otherwise C. pneumoniae infections were rarely seen in children. A Comparison of serology diagnostic tests for M. pneumoniae infections was found more sensitive and specific than PCR.

Auxo-, sero-, and opa-typing of Neisseria gonorrhoeae strains isolated in Istanbul, Turkey.

BACKGROUND: Typing methods are essential in understanding of the transmission dynamics of Neisseria gonorrhoeae. Several typing methods were described including opa-typing. GOALS: The goal of this study was to type all of the strains we isolated in the recent past by using auxo-, sero-, and opa-typing, and to compare the discriminatory power of these methods. STUDY: Auxotyping, serotyping, and opa-typing were performed for 56 N. gonorrhoeae strains isolated from male patients with urethritis. RESULTS: A total of 9 auxotypes and 33 serovars were detected. Combining the 2 systems, a total of 45 distinct auxotype/serovar (A/S) classes were identified. The most common A/S class was NR/Bsty (5 strains). Fifty-five distinct patterns were detected by opa-typing. Two strains that have been isolated 16 months apart gave identical patterns with opa-typing and their A/S class was also identical (NR/Bsty). Simpson's index of diversity was found as 0.664, 0.961, 0.987, and 0.999 for auxotyping, serotyping, A/S class, and opa-typing, respectively. CONCLUSIONS: Opa-typing is a potential useful method for typing N. gonorrhoeae as a result of its high discriminatory power, rapidity, ease and relatively lower cost.