The transcription factor PU.1 is required for the development of IL-9-producing T cells and allergic inflammation.

CD4(+) helper T cells acquire effector phenotypes that promote specialized inflammatory responses. We show that the ETS-family transcription factor PU.1 was required for the development of an interleukin 9 (IL-9)-secreting subset of helper T cells. Decreasing PU.1 expression either by conditional deletion in mouse T cells or the use of small interfering RNA in human T cells impaired IL-9 production, whereas ectopic PU.1 expression promoted IL-9 production. Mice with PU.1-deficient T cells developed normal T helper type 2 (T(H)2) responses in vivo but showed attenuated allergic pulmonary inflammation that corresponded to lower expression of Il9 and chemokines in peripheral T cells and in lungs than that of wild-type mice. Together our data suggest a critical role for PU.1 in generating the IL-9-producing (T(H)9) phenotype and in the development of allergic inflammation.

Interleukin-9 is required for allergic airway inflammation mediated by the cytokine TSLP.

Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine important for the initiation and development of T helper (Th2) cell-mediated allergic inflammation. In this study, we identified a positive association between interleukin-9 (IL-9) and TSLP concentration in the serum of infants with atopic dermatitis. In primary cell cultures, the addition of TSLP led to an increase in IL-9 production from human and mouse Th9 cells, and induced an increase in signal transducer and activator of transcription 5 (STAT5) activation and binding to the Il9 promoter. In vivo, use of an adoptive transfer model demonstrated that TSLP promoted IL-9-dependent, Th9 cell-induced allergic inflammation by acting directly on T cells. Moreover, transgenic expression of TSLP in the lung stimulated IL-9 production in vivo, and anti-IL-9 treatment attenuated TSLP-induced airway inflammation. Together, our results demonstrate that TSLP promotes Th9 cell differentiation and function and define a requirement for IL-9 in TSLP-induced allergic inflammation.

The ETS Family Transcription Factors Etv5 and PU.1 Function in Parallel To Promote Th9 Cell Development.

The IL-9-secreting Th9 subset of CD4 Th cells develop in response to an environment containing IL-4 and TGF-ß, promoting allergic disease, autoimmunity, and resistance to pathogens. We previously identified a requirement for the ETS family transcription factor PU.1 in Th9 development. In this report, we demonstrate that the ETS transcription factor ETS variant 5 (ETV5) promotes IL-9 production in Th9 cells by binding and recruiting histone acetyltransferases to the Il9 locus at sites distinct from PU.1. In cells that are deficient in both PU.1 and ETV5 there is lower IL-9 production than in cells lacking either factor alone. In vivo loss of PU.1 and ETV5 in T cells results in distinct effects on allergic inflammation in the lung, suggesting that these factors function in parallel. Together, these data define a role for ETV5 in Th9 development and extend the paradigm of related transcription factors having complementary functions during differentiation.

PU.1 Expression in T Follicular Helper Cells Limits CD40L-Dependent Germinal Center B Cell Development.

PU.1 is an ETS family transcription factor that is important for the development of multiple hematopoietic cell lineages. Previous work demonstrated a critical role for PU.1 in promoting Th9 development and in limiting Th2 cytokine production. Whether PU.1 has functions in other Th lineages is not clear. In this study, we examined the effects of ectopic expression of PU.1 in CD4(+) T cells and observed decreased expression of genes involved with the function of T follicular helper (Tfh) cells, including Il21 and Tnfsf5 (encoding CD40L). T cells from conditional mutant mice that lack expression of PU.1 in T cells (Sfpi1(lck-/-)) demonstrated increased production of CD40L and IL-21 in vitro. Following adjuvant-dependent or adjuvant-independent immunization, we observed that Sfpi1(lck-/-) mice had increased numbers of Tfh cells, increased germinal center B cells (GCB cells), and increased Ab production in vivo. This correlated with increased expression of IL-21 and CD40L in Tfh cells from Sfpi1(lck-/-) mice compared with control mice. Finally, although blockade of IL-21 did not affect GCB cells in Sfpi1(lck-/-) mice, anti-CD40L treatment of immunized Sfpi1(lck-/-) mice decreased GCB cell numbers and Ag-specific Ig concentrations. Together, these data indicate an inhibitory role for PU.1 in the function of Tfh cells, germinal centers, and Tfh-dependent humoral immunity.

STAT6-dependent regulation of Th9 development.

Th cell effector subsets develop in response to specific cytokine environments. The development of a particular cytokine-secreting pattern requires an integration of signals that may promote the development of opposing pathways. A recent example of this paradigm is the IL-9-secreting Th9 cell that develops in response to TGF-ß and IL-4, cytokines that, in isolation, promote the development of inducible regulatory T cells and Th2 cells, respectively. To determine how the balance of these factors results in priming for IL-9 secretion, we examined the effects of each pathway on transcription factors that regulate Th cell differentiation. We demonstrated that TGF-ß induces the PU.1-encoding Sfpi1 locus and that this is independent of IL-4-induced STAT6 activation. IL-4-activated STAT6 is required for repressing the expression of T-bet and Foxp3 in Th9 cells, transcription factors that inhibit IL-9 production, and STAT6 is required for the induction of IRF4, which promotes Th9 development. These data established a transcription factor network that regulates IL-9 and demonstrated how combinations of cytokine signals generate cytokine-secreting potential by altering the expression of a panel of transcription factors.

Carving a Path to the Brain: A Study on Neurosurgery Career Choices.

BACKGROUND: Medical students often face challenges in choosing a career path due to limited exposure to specialized fields like neurosurgery. Understanding their perceptions and experiences is crucial in addressing the gaps in neurosurgical education and inspiring future neurosurgeons. METHODS: A cross-sectional study was conducted involving 461 medical students, utilizing convenience sampling. Data collection employed a validated, self-administered tool. Statistical analysis in SPSS Version 25 included t-tests and chi-square tests, comparing scores based on age, gender, year of study, and exposure to the formal neurosurgical rotations in their institute. Significance value was set at P < 0.05. RESULTS: In the study of 461 medical students, 79.8% identified with the 19-23 age group, and 63.8% affirmed neurosurgery exposure. Medical students' perceptions included: 167 (36.3%) students found neurosurgery teaching sufficient; 164 (35.6%) disagreed that obtaining neurosurgical history is difficult; 224 (48.6%) agreed on neurosurgical disease complexity; and 250 (54.2%) found these diseases challenging and interesting. A majority of 183 (39.7%) respondents agreed that neurosurgical diseases had poor outcomes. Regarding training for neurosurgical surgery, 205 (44.5%) participants strongly agreed on its length, and 215 (46.7%) consented to extensive operating hours. However, 167 (36.3%) strongly disagreed about the ample job prospects in Pakistan. CONCLUSIONS: Enhancing neurosurgery education with quality, consistency, and adaptability is essential to bridge gaps and inspire future neurosurgeons. These findings guide improvements in educational programs, preparing a skilled workforce to meet evolving health-care demands.

PU.1 regulates TCR expression by modulating GATA-3 activity.

The Ets transcription factor PU.1 is a master regulator for the development of multiple lineages during hematopoiesis. The expression pattern of PU.1 is dynamically regulated during early T lineage development in the thymus. We previously revealed that PU.1 delineates heterogeneity of effector Th2 populations. In this study, we further define the function of PU.1 on the Th2 phenotype using mice that specifically lack PU.1 in T cells using an lck-Cre transgene with a conditional Sfpi1 allele (Sfpi1(lck-/-)). Although deletion of PU.1 by the lck-Cre transgene does not affect T cell development, Sfpi1(lck-/-) T cells have a lower activation threshold than wild-type T cells. When TCR engagement is limiting, Sfpi1(lck-/-) T cells cultured in Th2 polarizing conditions secrete higher levels of Th2 cytokines and have greater cytokine homogeneity than wild-type cells. We show that PU.1 modulates the levels of TCR expression in CD4(+) T cells by regulating the DNA-binding activity of GATA-3 and limiting GATA-3 regulation of TCR gene expression. GATA-3-dependent regulation of TCR expression is also observed in Th1 and Th2 cells. In CD4(+) T cells, PU.1 expression segregates into subpopulations of cells that have lower levels of surface TCR, suggesting that PU.1 contributes to the heterogeneity of TCR expression. Thus, we have identified a mechanism whereby increased GATA-3 function in the absence of the antagonizing activity of PU.1 leads to increased TCR expression, a reduced activation threshold, and increased homogeneity in Th2 populations.

Antibodies and Fab fragments protect Cu,Zn-SOD against methylglyoxal-induced inactivation.

Methyl glyoxal (MG) is a highly reactive alpha-oxoaldehyde that plays an important role in non-enzymatic glycosylation reactions, formation of Advanced Glycation End products (AGEs) and other complications associated with hyperglycemia and related disorders. Unlike sugars, glycation by MG is predominantly arginine directed, which is particularly more damaging since arginine residues have a high-frequency occurrence in ligand and substrate recognition sites in receptor and enzyme active sites. Using bovine erythrocyte Cu,Zn-superoxide dismutase (SOD) as model enzyme, the potential of anti-enzyme antibodies in imparting protection against MG-induced inactivation was investigated. A concentration- and time-dependent inactivation of SOD was observed when the enzyme was incubated with MG. The enzyme lost over 80% activity on incubation with 5 mM MG for 5 days. More marked inactivation was observed in 24 h when the MG concentration was raised up to 30 mM. The SOD inactivation was accompanied by the formation of high molecular weight aggregates as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI/TOF mass spectrometry). Inclusion of specific anti-SOD antibodies raised in rabbits or monomeric Fab fragments derived thereof offered remarkable protection against MG-induced loss in enzyme activity. The protection, however, decreased with increase in the concentration of MG. SELDI/TOF mass spectrometry also revealed that the antibodies restricted the formation of high molecular weight aggregates. The results emphasize the potential of antibody based therapy in combating glycation and related complications.

Inactivation and modification of superoxide dismutase by glyoxal: prevention by antibodies.

Glyoxal is an endogenous compound, the levels of which are increased in various pathologies associated with hyperglycaemia and other related disorders. It has been reported to inactivate critical cellular enzymes by promoting their cross-linking and perpetuates advanced glycation end-product (AGE) formation. In this study, we used superoxide dismutase (SOD) as a model to investigate the ability of specific anti-enzyme antibodies and monomer Fab fragments to protect against glyoxal-induced deactivation and aggregate formation. We found that glyoxal deactivated SOD, in a concentration and time-dependent fashion. The enzymatic activity was monitored spectrophotometrically and it was found that enzyme lost approximately 95% of its original activity, when exposed to 10 mM glyoxal for 120 h. SDS-polyacrylamide gel electrophoresis demonstrated the formation of high molecular weight aggregates in SOD samples exposed to glyoxal. Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) showed increase in relative molecular mass (M(r)), upon exposure to glyoxal. Specific anti-enzyme antibodies and monomer Fab fragments markedly inhibited SOD deactivation caused by glyoxal and decreased the extent of cross-linking or formation of aggregates. This protection by the antibodies or Fab fragments was specific since, other non-specific antibodies were not able to protect SOD. Previously, antibodies have been used to prevent aggregation of beta-amyloid peptides in Alzheimer and prion-protein disease. Our findings provide a new perspective, for use of antibodies to prevent the biomolecules against glycation-induced deactivation and alteration.

Retroviral Transduction and Reporter Assay: Transcription Factor Cooperation in Th9 Cell Development.

Naïve CD4+ T cells differentiate into different T helper subsets in response to specific cytokine environment and transcription factors. Th9 cells are induced in response to signals from cytokines, TGF-ß and IL-4. Transcription factors that are downstream of these cytokines converge to drive the development of Th9 cells. Retroviral transduction allows the genetic modification in T cells thereby helping us to better understand the molecular mechanisms that control their development as well as function. In this chapter, an optimized protocol for retroviral transduction of murine Th9 cells as well as transient transfection of Th9 cells with luciferase reporter constructs is described.

Analytical evaluation of HgbA1c, microalbumin, CRP, and RF on Architect ci8200 integrated system and workflow performance evaluation using computer simulation.

BACKGROUND: Recently, hemoglobin A1c (HgbA1c), microalbumin (MA), C-reactive protein (CRP) and rheumatoid factor (RF) have been introduced on high throughput general chemistry system. We evaluated analytical performance of these assays on an integrated clinical chemistry and immunoassay analyzer and studied the impact of testing these assays on these systems on the overall efficiency of the analyzer, via computer simulation. METHODS: The analytical performance was measured by determining precision, linearity and correlation of patient sample results with in-house testing methodology. MedModel simulation software is used to develop simulation model and process efficiency is determined by measuring turnaround times and resource utilization. RESULTS: Between-days CVs ranged from 8.59% for MA to 3.22% for HgbA1c level 1 controls. Less than 2% carryover for all 4 methods was observed on the integrated analyzer. For HgbA1c on HPLC analyzer, the minimum and maximum TAT for a batch of 50 samples was 3.78 and 160 min, respectively, while for the integrated system it was 28.2 and 35.1 min, respectively. Labor utilization for the 2 processes ranged from 3.21% to 3.75%. CONCLUSION: Chemistry module on an integrated system can be used to determine the HgbA1c and other serum proteins.

Ethnomedicinal uses of plants in the treatment of paediatric geohelminth infections in Kalat district of Northern Balochistan, Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: Infection by intestinal parasitic worms (soil-transmitted helminths or geohelminths) is prevalent in many parts of the world, and poses a particular health risk to children. This paper presents findings from a preliminary study with the primary aim to document indigenous knowledge about the use of herbal medicines in the treatment of intestinal worm infections in children among the local communities of Kalat district of northern Balochistan, Pakistan. MATERIALS AND METHODS: Ethnomedicinal data were collected through a triangulation approach, that included participant-observation and rapid appraisal methods. Prior-informed consent (PIC) was obtained from participants before conducting structured and semi-structured interviews and delivering an open ended questionnaire. A total of 94 participants, including 28 men (of whom 7 were traditional healers), and 66 women of four different age groups were interviewed. Results were analyzed using quantitative indices of Use Value citations (UVC) and Disease-Consensus Index (DCI). RESULTS: Fewer men than women agreed to be interviewed, thus overall women in the area appeared to have more ethnomedicinal knowledge. The majority of study participants belonged to the older age group (>55 years). A total of 49 plant species, belonging to 47 genera, distributed in 30 families were reported. The families Asteraceae and Lamiaceae were most frequently represented, with four species each. Trees were the most common life form, with seeds the most frequently cited plant part used (29%). Nearly a third (31%) of plant-based remedies reported in the treatment of intestinal worms were administered as a decoction. The highest UVC and DCI was reported for the species Ferula assa-foetida sL. (UVC 0.51, DCI 0.46). CONCLUSIONS: This study provides previously unreported data on the use of medicinal plants in the treatment of geohelminth infections in children of Kalat. Eight species, Acacia modesta Wall., Asparagus capitatus Baker, Microcephala lamellata (Bunge) Pobed., Nepeta praetervisa Rech.f., Plantago ciliata Desf., Pistacia atlantica Desf., Seriphidium quettense (Podlech) Y.R.Ling and Thymus linearis Benth. are reported here as anthelmintics for the first time. Detailed studies on the anthelmintic activity of chemical constituents of these species are lacking from existing literature. Further phytochemical, pharmacological and toxicity studies are required in order to evaluate the efficacy and safety of these newly reported anthelmintic species. These plants may provide a source of novel anthelmintic drug leads, which are urgently required due to the problem of global anthelmintic resistance.

Altered STAT4 Isoform Expression in Patients with Inflammatory Bowel Disease.

BACKGROUND: Crohn's disease (CD) and ulcerative colitis (UC) are the major forms of inflammatory bowel disease, and pathogenesis involves a complex interplay among genetic, environmental, and immunological factors. We evaluated isoform expression of the IL-12-activated transcription factor STAT4 in children with CD and UC. METHODS: We collected biopsy samples from both patients newly diagnosed with CD and with UC. We further collected blood samples from patients newly diagnosed with CD and with UC as well as from patients who had a flare-up after being in clinical remission, and we examined the ratios of STAT4ß/STAT4α mRNA. In addition to STAT4 isoforms, we measured the expression of the cytokines TNFα, IFNγ, granulocyte macrophage-colony stimulating factor, and IL-17 using polymerase chain reaction of biopsy samples and multiplex analysis of patient serum samples. RESULTS: Ratios of STAT4ß/STAT4α were increased in specific gastrointestinal tract segments in both patients with CD and those with UC that correlate with the location and severity of inflammation. In contrast, we did not observe changes in STAT4ß/STAT4α ratios in biopsy specimens from patients with eosinophilic esophagitis. We also observed increased STAT4ß/STAT4α ratios in the peripheral blood mononuclear cells of patients with UC and those with CD, compared with healthy controls. Ratios were normalized after patients were treated with steroids. CONCLUSIONS: Collectively, these data indicate that STAT4 isoforms could be an important noninvasive biomarker in the diagnosis and treatment of inflammatory bowel disease and that expression of these isoforms might provide further insight into the pathogenesis of IBD.

STAT4 deficiency reduces the development of atherosclerosis in mice.

Atherosclerosis is a chronic inflammatory process that leads to plaque formation in large and medium sized vessels. T helper 1 (Th1) cells constitute the majority of plaque infiltrating pro-atherogenic T cells and are induced via IFNγ-dependent activation of T-box (Tbet) and/or IL-12-dependent activation of signal transducer and activator of transcription 4 (STAT4). We thus aimed to define a role for STAT4 in atherosclerosis. STAT4-deficiency resulted in a ∼71% reduction (p < 0.001) in plaque burden in Stat4(-/-)Apoe(-/-) vs Apoe(-/-) mice fed chow diet and significantly attenuated atherosclerosis (∼31%, p < 0.01) in western diet fed Stat4(-/-)Apoe(-/-) mice. Surprisingly, reduced atherogenesis in Stat4(-/-)Apoe(-/-) mice was not due to attenuated IFNγ production in vivo by Th1 cells, suggesting an at least partially IFNγ-independent pro-atherogenic role of STAT4. STAT4 is expressed in T cells, but also detected in macrophages (MΦs). Stat4(-/-)Apoe(-/-)in vitro differentiated M1 or M2 MΦs had reduced cytokine production compare to Apoe(-/-) M1 and M2 MΦs that was accompanied by reduced induction of CD69, I-A(b), and CD86 in response to LPS stimulation. Stat4(-/-)Apoe(-/-) MΦs expressed attenuated levels of CCR2 and demonstrated reduced migration toward CCL2 in a transwell assay. Importantly, the percentage of aortic CD11b(+)F4/80(+)Ly6C(hi) MΦs was reduced in Stat4(-/-)Apoe(-/-) vs Apoe(-/-) mice. Thus, this study identifies for the first time a pro-atherogenic role of STAT4 that is at least partially independent of Th1 cell-derived IFNγ, and primarily involving the modulation of MΦ responses.

The endemic medicinal plants of Northern Balochistan, Pakistan and their uses in traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: The highlands of Northern Balochistan are the hot spots of medicinal and endemic plant in Pakistan. These plants are still commonly used for medicinal purposes by local people in their daily lives. This study first documented the information about the medicinal uses of endemic species of Balochistan-province Pakistan. MATERIALS AND METHODS: A survey was performed using open ended questionnaires, free listening and personal observations with 152 informants (54% female, 46% male). In addition, the use value (MUV), use report (UR), fidelity level (FL), frequency citation (FC), relative frequency citation (RFC), family importance value (FIV) of species were determined and the informant consensus factor (ICF) was calculated for the medicinal plants included in the study. RESULTS: A total of 24 endemic plants belonging to 19 genera and 14 families were used by the local inhabitants to treat 12 categories of various diseases. The most common families of endemic plant species as depicted by its number of species (6 species) and FIV (9.9) was Fabaceae as the dominant family. The endemic plant species comprised perennial herbs (30%), annual herbs (25%), shrubs (29%) and under shrubs (16% each), no endemic tree species was reported in the study area. The highest number of species were used in the treatment of gastrointestinal diseases (12 species). The main route of administration is oral injection (62%) while the most frequently used form of external administration of herbal medicine was paste (5.4%) and the most commonly applied methods of preparation are powder (48.2%). Highest use report were calculated for Allium baluchistanicum and Viola makranica, (8 UR each), and least use report were calculated for two species Heliotropium remotiflorum and Tetracme stocksii (1 UR for each). Use values of the recorded plant species have been calculated which showed a highest use value of (0.73) for A. baluchistanicum and (0.56) for Berberis baluchistanica while the lowest UVs were attained for T. stocksii (0.13). Highest RFC value were calculated for Achillea millefolium (0.19) and least RFC were calculated for Blepharis sindica (0.02). The endemic species with 100% fidelity level was calculated for two plant species i.e. Seriphidium quettense and B. baluchistanica. CONCLUSIONS: The Balochistan is rich in endemic and other medicinal plants, still needs more exploration and study. Thus, it is important to document and reconstitute the remainders of the ancient medical practices which exist in Balochistan and other areas of the world, and preserve this knowledge for future generations. The endemic species which are used in traditional medicine in the region lacks phototherapeutic evidence. It is necessary to perform phytochemical or pharmacological studies to explore the potential of plants used for medicinal purposes. Overgrazing, urbanization and unsustainable harvesting of such rare and endemic medicinal plants in this region is facing severe threats of extinction. It is thus recommended that cultivation techniques be formulated, especially for the most important endemic plant medicinal species of the region.

Ethnobotany of medicinal plants in district Mastung of Balochistan province-Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: The aim of this study was to document the medicinal uses of plants in district Mastung of Balochistan province, Pakistan. The ethnobotanical results contain quantitative information on medicinal plants diversity documented for the first time in the area. MATERIALS AND METHODS: The information was collected through semi-structured interviews, rapid appraisal approach, open ended questionnaire and personal observations. Results were analyzed using quantitative indices of information consent factor (ICF), fidelity level (FL), use value (UV), frequency citation (FC) and relative frequency citation (RFC). RESULTS: In total of 102 plant species belonging to 47 families were reported for the medicinal purposes. Asteraceae was found to be dominant family in terms of species in the area with 11 species. The whole plant and leaves were noted as most frequently used parts (24%). Decoction (31% with 40 species) was the most commonly used preparation method. Highest ICF value (1) was recorded for antidote category. 100% fidelity level was found for four plant species i.e. Achillea welhemsii, Caralluma tuberculata, Citrullus colocynthis, and Seripidium quettense. The highest use value was reported for the Acroptilon repens (0.5) while highest RFC value was calculated for Berberis balochistanica and Citrullus colocynthis (0.18). Highest use report was calculated for Caralluma tuberculata, Citrullus colocynthis, Malva neglecta and Mentha longifolia with five use reports for each. CONCLUSIONS: The area is rich in medicinal plants and these plants are still commonly used for medicinal purposes among the people in their daily lives. However, there is a gradual loss of traditional knowledge about these plants in new generation. This study provides basis for the conservation of the local flora, its use as food and medicine. It also provides various socio-economic dimensions associated with the common people.

Th9 cell development requires a BATF-regulated transcriptional network.

T helper 9 (Th9) cells are specialized for the production of IL-9, promote allergic inflammation in mice, and are associated with allergic disease in humans. It has not been determined whether Th9 cells express a characteristic transcriptional signature. In this study, we performed microarray analysis to identify genes enriched in Th9 cells compared with other Th subsets. This analysis defined a transcriptional regulatory network required for the expression of a subset of Th9-enriched genes. The activator protein 1 (AP1) family transcription factor BATF (B cell, activating transcription factor­like) was among the genes enriched in Th9 cells and was required for the expression of IL-9 and other Th9-associated genes in both human and mouse T cells. The expression of BATF was increased in Th9 cultures derived from atopic infants compared with Th9 cultures from control infants. T cells deficient in BATF expression had a diminished capacity to promote allergic inflammation compared with wild-type controls. Moreover, mouse Th9 cells ectopically expressing BATF were more efficient at promoting allergic inflammation than control transduced cells. These data indicate that BATF is a central regulator of the Th9 phenotype and contributes to the development of allergic inflammation.

The symphony of the ninth: the development and function of Th9 cells.

CD4+ T helper cells are obligate regulators of inflammatory disease. An expanding cadre of T helper (Th) subsets, specialized for promoting particular types of inflammation, function through the secretion of a restricted set of cytokines. The latest addition to the list of subsets is the Th9 cell that secretes IL-9 as a signature cytokine and contributes to several classes of inflammatory disease. In this review we focus on recent advances in understanding the development of Th9 cells, and how Th9 cells contribute to the orchestration of disease.

The transcription factor PU.1 regulates γδ T cell homeostasis.

BACKGROUND: T cell development results in the generation of both mature αß and γδ T cells. While αß T cells predominate in secondary lymphoid organs, γδ T cells are more abundant in mucosal tissues. PU.1, an Ets family transcription factor, also identified as the spleen focus forming virus proviral integration site-1 (Sfpi1) is essential for early stages of T cell development, but is down regulated during the DN T-cell stage. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we show that in mice specifically lacking PU.1 in T cells using an lck-Cre transgene with a conditional Sfpi1 allele (Sfpi1(lck-/-)) there are increased numbers of γδ T cells in spleen, thymus and in the intestine when compared to wild-type mice. The increase in γδ T cell numbers in PU.1-deficient mice is consistent in γδ T cell subsets identified by TCR variable regions. PU.1-deficient γδ T cells demonstrate greater proliferation in vivo and in vitro. CONCLUSIONS/SIGNIFICANCE: The increase of γδ T cell numbers in Lck-Cre deleter strains, where deletion occurs after PU.1 expression is diminished, as well as the observation that PU.1-deficient γδ T cells have greater proliferative responses than wild type cells, suggests that PU.1 effects are not developmental but rather at the level of homeostasis. Thus, our data shows that PU.1 has a negative influence on γδ T cell expansion.

Polyclonal antibodies inhibit the glycation-induced inactivation of bovine Cu,Zn-superoxide dismutase.

In vitro incubation of bovine Cu,Zn-SOD (Cu,Zn-superoxide dismutase) with glucose, ribose or fructose results in a remarkable inactivation of the enzyme. There was a progressive decrease in enzyme activity on incubation with glucose and, at the end of 7 days, only 26% of the initial activity remained. The inactivation was accompanied by a parallel decrease in the amount of protein detectable on gels after SDS/PAGE. Reaction of the sugars with SOD was ascertained using an immunoblot assay in which sugar-incubated SOD was derivatized with 2,4-dinitrophenylhydrazine and allowed to react with the dinitrophenol-specific antibody. Affinity purified antibodies from the sera of rabbits immunized with bovine SOD were highly effective in restricting the inactivation of the enzyme induced by glucose, ribose or fructose.