TNF-α - 308 G/A and IFN-γ + 874 A/T gene polymorphisms in Saudi patients with cutaneous leishmaniasis.

BACKGROUND: Cutaneous leishmaniasis (CL) is well linked with immunogenetic factors. This study was undertaken to test the association of TNF-α - 308 and IFN-γ + 874 gene polymorphisms with the susceptibility of Leishmania (L) species among CL patients in central region of Saudi Arabia. METHODS: This is a case-control study involved 169 Saudi subjects with different L. species and 199 healthy controls from central region of Saudi Arabia. All subjects were characterized by TNF-α - 308 G/A and IFN-γ + 874 A/T gene polymorphisms using PCR. RESULTS: Evaluation of genotyping and allelic frequency of TNF-α - 308 G/A in different L. species showed no significant association compared to controls (p > 0.05). Except, in cases of L. tropica that showed significantly higher TNF-α - 308 A versus G allele frequency (p = 0.0004). Evaluation of genotyping of IFN-γ + 874 (TT versus AA+AT recessive) and allelic frequency of IFN-γ + 874 (T versus A) showed significant higher in L. major and also in total CL cases as compared to healthy controls (p < 0.05). Furthermore, a strong association was observed between the susceptibility of L. major, L. tropica or total CL cases with synergistically combined high TNF-α 308/INF-γ 874 alleles. CONCLUSIONS: This is the first report that shows the gene polymorphisms of TNF-α - 308 G/A and IFN-γ + 874 A/T in Saudi patients with different L. species infections. Data showed that the TNF-α-308 G/A gene polymorphism is not associated with the susceptibility of CL in Saudi subjects. The only correlation was found in between A versus G allelic frequency in L. tropica. Importantly, IFN-γ + 874 A/T polymorphism was found to be associated with the susceptibility of L. major and also with total CL subjects. Moreover, data from synergistically combined high TNF-α 308/INF-γ 874 alleles strongly suggest their potential role in the susceptibility of leishmania infection.

Prevalence of Leishmania species among patients with cutaneous leishmaniasis in Qassim province of Saudi Arabia.

BACKGROUND: Leishmaniasis is a parasitic infection endemic in more than ninety countries of the world. The cutaneous leishmaniasis (CL) is a most common form of leishmaniasis and it remains to be a major public health issue in Saudi Arabia. This study was undertaken to investigate the Leishmania species responsible for CL infection in different provinces of Qassim, Saudi Arabia. METHODS: Skin biopsies were obtained from CL patients and DNA was extracted using the Magna pure system. Leishmania species were identified by highly specific/sensitive quantitative and qualitative PCR. RESULTS: Out of total 206 CL biopsies, 49.5% biopsies were found to be positive for Leishmania major (L. major), 28.6% biopsies were positive for Leishmania tropica (L. tropica), 3.9% were found to be positive for Leishmania infantum/donovani (L. infantum/donovani). Not only have these, all tested CL biopsies showed negative test for Leishmania mexicana (L. mexicana) and Leishmania viannia (L. viannia). CONCLUSIONS: This is the first comprehensive study that shows the majority of CL in Qassim was caused by L. major and L. tropica. To the best of our knowledge, this is the very first report that shows the occurrence of L. infantum/donovani in Saudi Arabia. This requires higher alert to the Ministry of Health of Saudi Arabia to take proactive actions in preventing the onset of L. major, L. tropica, L. infantum and L. donovani infections.

Markers of atopic dermatitis, allergic rhinitis and bronchial asthma in pediatric patients: correlation with filaggrin, eosinophil major basic protein and immunoglobulin E.

BACKGROUND: Allergic reactions have been implicated as contributions in a number of atopic disorders, including atopic dermatitis (AD), allergic rhinitis (AR) and bronchial asthma (BA). However, the potential for filaggrin protein, eosinophil major basic protein (MBP) and immunoglobulin E (IgE) to elicit allergic response or to contribute to atopic disorders remains largely unexplored in pediatric patients. This study was undertaken to investigate the status and contribution of filaggrin protein, eosinophil MBP and total IgE in pediatric patients with AD, AR and BA. METHODS: Sera from 395 pediatric patients of AD, AR or BA with varying levels of disease activity according to the disease activity index and 410 age-matched non-atopic healthy controls were evaluated for serum levels of atopic markers, including filaggrin, eosinophil MBP and IgE. RESULTS: Serum analysis showed that filaggrin levels were remarkably high in pediatric patients with AD, followed by BA and AR, whereas its levels were low in non-atopic pediatric controls. Eosinophil MBP levels in sera of atopic patients were significantly high as compared with their respective controls, but its levels were highest in AR patients, followed by AD and BA. Total IgE in sera of AD patients was markedly high, followed by AR and BA patients, whereas its levels were low in non-atopic pediatric controls. Interestingly, not only was an increased number of subjects positive for filaggrin protein, eosinophil MBP or total IgE, but also their levels were statistically significantly higher among those atopic patients whose disease activity scores were higher as compared with atopic patients with lower disease activity scores. CONCLUSIONS: These findings strongly support a role of filaggrin protein, eosinophil MBP and IgE in the onset of allergic reactions in pediatric patients with AD, AR and BA. The data suggest that filaggrin, eosinophil MBP or IgE might be useful in evaluating the progression of AD, AR or BA and in elucidating the mechanisms involved in the pathogenesis of these pediatric disorders.

Acquired immunogenicity of DNA after modification with malondialdehyde in patients with alopecia areata.

BACKGROUND: Lipid peroxidative-mediated reactions have been implicated in alopecia areata (AA). However, the potential for lipid peroxidation to elicit an autoimmune response or to contribute in disease pathogenesis remain unexplored. This study was undertaken to investigate the status/contribution of lipid oxidative-by-product malondialdehyde modified DNA (MDA-DNA) in AA. METHODS: DNA was modified by MDA. Binding characteristics of AA circulating immunoglobulin G (AA-IgG) MDA-modified DNA were screened. Immunogenicity of MDA-DNA was probed by inducing antibodies in rabbits. DNA samples from AA patients were isolated (DNA-AA) and their immune reactions with rabbit anti-MDA-DNA-IgGs were evaluated. RESULTS: Our data show that MDA caused extensive DNA damage. Protein-A purified IgG from 45% of AA patients showed strong binding to MDA-DNA in comparison with native DNA (p < 0.05). MDA-DNA was found to be highly immunogenic in rabbits. Rabbit anti-MDA-DNA-IgG showed binding with isolated DNA from AA patients. CONCLUSIONS: This is the first study to demonstrate the role of MDA-modified DNA in AA patients. Our novel results conclude that perturbations in DNA by MDA present unique neo-epitopes that might be one of the factors for the antigen driven antibodies induction in AA. These results provide an important insight into the immunological mechanisms occurring in AA.

4-Hydroxy-2-nonenal modified histone-H2A: a possible antigenic stimulus for systemic lupus erythematosus autoantibodies.

Protein modifications by 4-hydroxy-2-nonenals (HNE) are involved in various diseases. Histones are DNA protective nucleoprotein, which adopt different structures under oxidative stress. This study was undertaken to test the role of HNE-modified-histone-H2A (HNE-H2A) in systemic lupus erythematosus (SLE). Our data revealed that HNE-mediated-lipid peroxidation in histone-H2A caused alteration in histidine, lysine and cystein residues. In addition, protein carbonyl contents were also high in HNE-H2A. HNE-specific quencher, L-carnosine further reiterates HNE-modifications. Specificity of autoantibodies from SLE patients (n=48) were analyzed towards HNE-H2A and their results were compared with sex- and age-matched controls (n=36). SLE autoantibodies show preferential binding to HNE-H2A in comparison with histone-H2A (p<0.0001). Furthermore, HNE-H2A was also detected in SLE peripheral blood mononuclear cells. In conclusion, this is the first study to demonstrate the role of HNE-modified-histone in SLE. Preferential binding of HNE-H2A by affinity purified SLE-IgG pointed out the likely role of HNE-H2A in the initiation/progression of SLE.

Immunological studies of reactive oxygen species damaged catalase in patients with systemic lupus erythematosus: correlation with disease activity index.

OBJECTIVES: This study was undertaken to investigate the status and contribution of oxidized catalase in systemic lupus erythematosus (SLE) and to explore whether oxidized catalase has a role in disease progression. METHODS: Catalase (CAT) was modified by reactive oxygen species (ROS). Sera from 50 SLE patients with varying levels of disease activity according to SLE Disease-Activity-Index (SLEDAI) and 45 age- and sex-matched healthy controls were evaluated for antibodies against oxidized CAT. RESULTS: Serum analysis showed significantly higher level of anti-oxidized-CAT-antibodies in SLE patients compared with controls. Interestingly, not only was there an increased number of subjects positive for anti-oxidized-CAT-antibodies, but also the levels of these antibodies were significantly higher among SLE patients, whose SLEDAI scores were ≥ 10 as compared with lower SLEDAI scores (<10). In addition, significant correlation was observed between the levels of anti-oxidized-CAT-antibodies and SLEDAI score (r = 0.796). Furthermore, sera from SLE patients had lower levels of CAT activity compared with control sera. CONCLUSIONS: These findings support an association between oxidized CAT and SLE. The stronger response observed in serum samples from patients with higher SLEDAI scores suggests that oxidized CAT may be a useful biomarker in evaluating the progression of SLE and in elucidating the mechanisms of disease pathogenesis.

Biochemical markers of oxidative and nitrosative stress in acne vulgaris: correlation with disease activity.

BACKGROUND: Acne vulgaris is a multifactorial skin disorder of unknown etiology. Free radical-mediated reactions have been implicated but their role in eliciting this response and contributing to disease progress remains unexplored. This study was undertaken to investigate the status and contribution of oxidative/nitrosative stress in patients with acne vulgaris. METHODS: Sera from 50 acne vulgaris with varying levels of disease activity (mild, moderate, and severe) according to the Global Acne Grading System (GAGS) and 40 age- and sex-matched controls were evaluated for serum levels of oxidative/nitrosative stress markers, including protein oxidation, lipid peroxidation and nitric oxide (NO), superoxide dismutase (SOD), and glutathione (GSH). RESULTS: Serum analysis showed significantly higher levels of carbonyl contents, malondialdehyde (MDA) and NO, in acne patients compared with healthy controls (P < 0.05). Interestingly, not only there were an increased number of subjects positive for carbonyl contents, but also the levels of these oxidants were significantly increased with the increase of the disease activity (P < 0.05). In addition, a significant correlation was observed between the levels of carbonyl contents and the GAGS scores (r = 0.341, r = 0.355, and r = 0.299, respectively). Furthermore, sera from acne patients had lower levels of SOD and GSH compared with healthy control sera. CONCLUSION: These findings support an association between oxidative/nitrosative stress and acne. The stronger response observed in serum samples from patients with higher GAGS scores suggests that markers of oxidative/nitrosative stress may be useful in evaluating the progression of acne and in elucidating the mechanisms of disease pathogenesis.

Macrolides in chronic inflammatory skin disorders.

Long-term therapy with the macrolide antibiotic erythromycin was shown to alter the clinical course of diffuse panbronchiolitis in the late 1980s. Since that time, macrolides have been found to have a large number of anti-inflammatory properties in addition to being antimicrobials. These observations provided the rationale for many studies performed to assess the usefulness of macrolides in other inflammatory diseases including skin and hair disorders, such as rosacea, psoriasis, pityriasis rosea, alopecia areata, bullous pemphigoid, and pityriasis lichenoides. This paper summarizes a collection of clinical studies and case reports dealing with the potential benefits of macrolides antibiotics in the treatment of selected dermatoses which have primarily been classified as noninfectious and demonstrating their potential for being disease-modifying agents.

MicroRNA-183-5p regulates MITF expression in vitiligo skin depigmentation.

Microphthalmia-associated transcription factor (MITF) is a master regulatory factor for melanocytes. MITF regulates multiple pigmentary genes for maintaining cellular homeostasis. MicroRNAs (miRNAs) play crucial roles in numerous biological processes however their molecular/cellular mechanisms to regulate pigmentation have not been fully explored. This study was undertaken to investigate the role of miRNAs in skin depigmentation via regulation of MITF gene. Depigmentation in C57BL/6 black mice was induced by an autoimmune response against tyrosinase. Bioinformatics approach was used to detect miRNAs conserved in 3'untraslated region (3'UTR) of MITF mRNA. The iMC23 mouse melanocytes were used for transfection experiments. The data demonstrated that the MITF mRNA/protein was markedly low in lesional skin of depigmented mice (p < 0.05). Targetscan genomic database determined that 3'UTR of mouse MITF constitutes 4819 nucleotide bases and has 23 conserved sites for different miRNAs To validate the pairing of these predicted miRNAs with MITF mRNA, five miRNAs were deregulated in lesional skin (p < 0.05). Among them, mmu-miR-181a-5p and mmu-miR-183-5p were up-regulated, whereas mmu-miR-26a-5p, mmu-miR-26b-5p and mmu-miR-32-5p were down-regulated (p < 0.05). To verify these results, the iMC23 mouse melanocytes were used. Transfection of iMC23 cells with specific miRNAs mimics or inhibitors or with 3'UTR reporter clone of MITF, showed only mmu-miR-183-5p binds to 3'UTR of MITF mRNA and regulates its expression in iMC23 melanocytes. In conclusions, this is the first study shows that miR-183-5p is a direct regulator of MITF in iMC23 melanocytes. Thus, miR-183-5p is an important regulator of melanocytes homeostasis and may be a novel target for autoimmune depigmentation therapy.

Obstetrical and fetal outcomes of a new management strategy in patients with intra-hepatic cholestasis of pregnancy.

PURPOSES: To determine the incidence, obstetrical, and fetal complication rates of intrahepatic cholestasis of pregnancy (ICP) in patients managed expectantly to 40-weeks gestation. METHODS: In a prospective cohort study conducted between February 2008 and January 2010, a total of 21,960 pregnant women in Qassim Region of Saudi Arabia were screened for ICP using specific criteria for diagnosis. The course of pregnancy was monitored to 40-weeks gestation or spontaneous onset of labor, whichever comes first. The measured outcomes were compared with a cross-matched group of healthy pregnant women. Continuous variables were analyzed with t test, while χ(2) test was used for comparing percentages. RESULTS: The incidence of ICP was 0.35% (76/21,960). There was no significant difference between groups in gestational age at delivery, preterm labor, intrauterine fetal death, cesarean section, or respiratory distress syndrome. There was significantly higher intrapartum non-reassuring fetal heart rate patterns and meconium-stained amniotic fluid in ICP group (P < 0.01 and <0.0001, respectively). CONCLUSIONS: The incidence of ICP in this region is low compared to worldwide range. Expectant management to 40-weeks gestation is associated with obstetrical and fetal outcomes comparable to normal pregnancy; however, intrapartum fetal asphyxia is more likely.

Skin manifestations in diabetic patients attending a diabetic clinic in the Qassim region, Saudi Arabia.

OBJECTIVE: To investigate the prevalence of skin manifestations in diabetic patients attending a diabetic clinic in the Qassim region, Saudi Arabia. SUBJECTS AND METHODS: A prospective observational study was performed on 320 patients (174 males and 146 females) attending the diabetic clinic. A detailed dermatological examination was carried out by a consultant dermatologist and the cutaneous findings were recorded. RESULTS: The overall prevalence of skin manifestations was 91.2%. Cutaneous lesions were seen in 12 patients (34.3%) of type 1 diabetes mellitus (DM) and 280 (98.2%) of type 2 diabetics. There was a statistically significant difference (p < 0.001) in skin manifestations between type 1 and type 2 DM patients. For those patients having diabetes of less than 5 years' duration, the incidence of skin manifestations was 80.6%; for those having had diabetes for more than 5 years, the incidence was 98%. This difference was statistically significant (p < 0.001). The skin manifestations that had a statistically significant difference (p < 0.05) in prevalence between the 2 durational groups were gangrene, diabetic dermopathy, paresthesia, diabetic feet, diabetic bullae and fungal infections. CONCLUSION: Diabetics had a greater prevalence of skin manifestations in type 2 than type 1, and as the duration of diabetes increased, the likelihood of developing skin manifestations also increased. Early referral to the dermatologist may help to detect complications of the skin in diabetes at an early stage and may prevent disability caused by these complications.

Missense, silent, non-sense and frame-shift mutations in exon 3 of the filaggrin gene in patients with bronchial asthma, atopic dermatitis, allergic rhinitis and mixed atopy.

This study investigated the atopic march on the basis of genetics. This research detected 227 variants in the filaggrin gene (FLG gene). Missense, silent, non-sense, frame-shift and non-coding mutations were detected in exon 3 of the FLG gene in patients with bronchial asthma, atopic dermatitis, allergic rhinitis and mixed atopy. Missense mutation was detected at c.8343 G > C (p. Asp2781Glu) in all adult asthmatic and allergic rhinitis patients. Whereas, mutation at c.8360 C > T/A (p. Arg2787 His/Leu) was detected in all childhood asthmatic and mixed atopic patients. A non-coding mutation was detected at c.12365 in atopic dermatitis and bronchial asthma patients. Furthermore, DNA sequencing of asthmatic and mixed atopic patients showed missense mutations at c.6073 C > T (p. Gly2025Glu) and a silent mutation at c. 8341 G > A (p. Asp2781Asp).

Autoimmune response against tyrosinase induces depigmentation in C57BL/6 black mice.

Regulation of melanogenesis by tyrosinase has now become an attractive approach for treatment of vitiligo but still the role of tyrosinase in the induction of depigmentation remains largely unexplored. This study was explored the role of tyrosinase in the induction of autoimmune depigmentation in C57BL/6 mice. Depigmentation was induced in C57BL/6 mice by tyrosinase immunization. Induced depigmentation was characterized by visual detection and was verified by histopathological analysis of lesional and non-lesinal skin biopsies. Moreover, induced depigmentation was re-validated by gene expression analysis of vitiligo-relevant genes by Taqman assays. Immunization of C57BL/6 mice by tyrosinase induces depigmentation on hairs as well as on skin. Immunoassays with Protein A-purified immune IgGs showed high titre antibodies against tyrosinase. Histopathological analysis showed that the total melanocytes were depleted from the basal layer of the epidermis and also from the dermis of depigmented lesions. The gene expression of vitiligo-relevant genes TYRP1, DCT, MLANA, MCIR, POMC, FOXJ2, CSNK1G3, SOX10, PMEL and KIT was significantly low in lesional skin as compared with non-lesional skin (p < .05). In contrast, the mRNA expression of CASP3 and NFκB1 was significantly high in lesional skin of depigmented mice as compared with non-lesional skin (p < .05). Furthermore, involvement of cellular immunity in depigmentation was confirmed by the reduction of CD4+:CD8+ lymphocytes ratio. In conclusion, this study shows that the autoimmune response against tyrosinase induces depigmentation in black C57BL/6 mice. The data obtained from the lesional and non-lesional skin biopsies showed the same features as were reported in human vitiligo patients.

Folliculitis decalvans.

Folliculitis decalvans is a rare inflammatory scalp disorder. The present paper gives a practical approach to diagnosis and patient management and reviews possible pathogenetic factors and treatment options. Folliculitis decalvans is classified as primary neutrophilic cicatricial alopecia and predominantly occurs in middle-aged adults. Staphylococcus aureus and a deficient host immune response seem to play an important role in the development of this disfiguring scalp disease. Lesions occur mainly in the vertex and occipital area. Clinically, the lesions present with follicular pustules, lack of ostia, diffuse and perifollicular erythema, follicular tufting, and, oftentimes, hemorrhagic crusts and erosions. Histology displays a mainly neutrophilic inflammatory infiltrate in early lesions and additionally lymphocytes and plasma cells in advanced lesions. Treatment is focused on the eradication of S. aureus anti-inflammatory agents.

Lichen planopilaris.

Lichen planopilaris is a chronic scarring alopecia characterized by follicular hyperkeratosis, perifollicular erythema, and loss of follicular orifices. The scalp lesions may be single or multiple and commonly involve the vertex and parietal area. The hair follicles at the margin of the alopecic patches reveal perifollicular erythema. Anagen hairs can be pulled out easily in active lesions. Associated cutaneous, nail, and mucous membrane lichen planus may be present. Commonly encountered symptoms and signs are increased hair shedding, itching, scaling, burning, and tenderness. Differentiation from other cicatricial alopecia can be performed through meticulous evaluation of the clinical, histopathologic, and immunohistopathologic findings. Treatment strategies depend on the disease activity and physician expertise. Although there are no definitive curative modalities, some new discoveries and conceptual advances continue to broaden our treatment options of this complex condition.

Pseudopelade of Brocq.

Pseudopelade of Brocq (PPB) is a rare, idiopathic, slowly progressive hair disorder, resulting in cicatricial alopecia. It typically presents in Caucasian adult patients as small, smooth, flesh-toned and slightly depressed alopecic patches with irregular outlines. It primarily involves the parietal and vertex portions of the scalp with a chronic prolonged course. Controversial opinions still exist as to whether PPB is a single entity or an end stage of several cicatricial alopecic disorders. A practical approach to diagnosis of PPB and therapeutic update are discussed in this review.

Filaggrin, major basic protein and leukotriene B4: Biomarkers for adult patients of bronchial asthma, atopic dermatitis and allergic rhinitis.

Bronchial asthma (BA), atopic dermatitis (AD), and allergic rhinitis (AR) are well known atopic disorders with complex etiologies. This study was undertaken to investigate the role of filaggrin, eosinophil major basic protein (MBP) and leukotriene B4 (LTB4) in patients with BA, AD, and AR. Sera from 1,246 patients with different atopic disorders and 410 normal healthy controls were collected and were evaluated for filaggrin, MBP and LTB4 by specific sandwich ELISAs, whereas immunoglobulin E (IgE) was used as a positive control for atopic patients. Serum analysis showed that filaggrin levels were remarkably high in patients with AD and in patients with multiple (mixed) atopic disorders (p < 0.001), whereas its levels in BA and AR patients were low but much higher than in normal human sera (p < 0.01). MBP levels were also high in AR, BA and mixed atopic patients, whereas AD patients showed no increase of MBP (p > 0.05). In contrast, LTB4 level was found to be significantly low in all tested atopic patients groups as compared to the levels of LTB4 present in normal human sera (p < 0.001). In conclusion, these findings support an association between filaggrin, MBP or LTB4 and atopic disorders. Our data strongly suggest that filaggrin, MBP or LTB4 might be useful in elucidating the mechanisms involved in the pathogenesis of these atopic disorders.

Association of psychological stress with skin symptoms among medical students.

OBJECTIVES: To evaluate the association between psychological stress and skin symptoms among medical students.  Methods: A cross-sectional study was carried out between January and June 2015. Electronic survey consists of Perceived Stress Questionnaire (PSQ) and Self-Reported Skin Complaints Questionnaire were distributed to all 1435 undergraduate students at College of Medicine, King Saud University (KSU), Riyadh, Saudi Arabia. Results: Final analysis was performed on data from 529 (36.9%) students. Students were divided into three groups: least stressed students, n=135, PSQ index less than 0.39; highly stressed students, n=136, PSQ index greater than 0.61; and moderately stressed students, n=258. Older age, female gender, during exam weeks, and fourth and fifth years of medical school (all p less than 0.01) were associated with the highest perceived stress levels. When compared to least stressed students, highly stressed students suffered from more oily, waxy patches or flakes on scalp (p≤0.0001), dry/sore rash (p≤0.0001), warts (p≤0.0001), pimples (p≤0.0001), itchy skin (p≤0.0001), hands itchy rash (p≤0.0001), hair loss (p≤0.0001), pull-out own hair (p=0.008), scaly skin (p=0.012), troublesome sweating (p=0.016), nails biting (p=0.028), and other rashes on face (p= 0.028).  Conclusion: Various common skin conditions could appear in context of psychological stress among medical students.

Altered expression of intracellular Toll-like receptors in peripheral blood mononuclear cells from patients with alopecia areata.

BACKGROUND: Toll-like receptors (TLRs) are pattern-recognition-receptors that sense a variety of pathogens and initiation of innate and adaptive immune responses. This study was undertaken to investigate the expression of TLRs in peripheral blood-mononuclear cells (PBMCs) of AA patients and to determine whether TLR-mediated inflammatory signals are important for the perspective of AA management. METHODS: Gene expression of TLRs and T-helper (Th) type-1, Th-2, Th-17 and regulatory T-cell cytokines in PBMCs was quantified by TaqMan Assays. Production of these cytokines in serum samples was determined by sandwich ELISAs. RESULTS: All TLRs (TLRs 1-10) were expressed in PBMCs of AA patients. Importantly intracellular TLRs (TLRs 3, 7, 8 and 9) were significantly up-regulated in AA patients as compared with controls (p < 0.05). Interleukin (IL)-2, TNF-α, and IL-17A gene expression in patients' PBMCs and their secretion in patients' sera were significantly higher as compared with their respective controls (p < 0.05). Whereas, TGF-ß gene expression in patients' PBMCs and TGF-ß protein level in patients' sera were significantly lower as compared with their controls (p < 0.05). CONCLUSION: This is the first report that shows the comprehensive expression profile of TLRs in AA patients. We conclude that up-regulated expression of intracellular TLRs in PBMCs of AA patients may play an active role in abnormal regulation of Th-1, Th-17 and regulatory T-cell cytokines in alopecia areata. GENERAL SIGNIFICANCE: Targeting of TLRs and their associated inflammatory signaling will open new areas of research; this may lead to the development of novel therapeutic targets for the treatment of AA or other skin disorders.

Interleukin-4 and interferon-γ are possible allergic markers in pediatric patients with ß-lactam hypersensitivity.

BACKGROUND: ß-lactam agents are known to elicit T-cell-mediated immune responses that play a central role in the onset of allergic reactions, but the involvement of specific type of cytokines in drug allergy remains largely unexplored in humans. OBJECTIVES: This study was undertaken to investigate the role of cytokines involvement in pediatric patients with ß-lactam hypersensitivity and to determine whether involvement of cytokines in drug-mediated reactions are important for the perspective of allergic patient's management. METHODS: ß-lactam-induced hypersensitivity reactions in eighty pediatric patients were determined by clinical manifestations and skin prick or intradermal testing. Production of T-helper (Th) type-1 cytokine interferon (INF)-γ, Th-2 cytokine interleukin (IL)-4, regulatory T-cell cytokine IL-10, and other cytokines IL-6 and IL-12 were determined by sandwich ELISAs. RESULTS: Diagnosis of ß-lactam allergy was confirmed in 53 pediatric patients. IL-4 secretion in patients' sera was significantly higher as compared with healthy controls (P < 0.05). However, INF-γ level in patients' sera was significantly lower as compared with controls (P < 0.05). No significant alterations were found in the protein secretion of IL-10, IL-12, and IL-6 in allergic patients as compared with controls (P > 0.05). CONCLUSION: We conclude that IL-4 is specific marker for the diagnosis of ß-lactam-induced hypersensitivity. Moreover, IL-4 in combination with INF-γ is more sensitive for the diagnosis of these reactions. This study also concludes that both IL-4 and INF-γ may play an active role in the onset of allergic reactions against ß-lactam antibiotics.