bluuebunny/crossref_metadata_2025_split huggingface dataset:
Th1 differentiation drives the accumulation of intravascular, non-protective CD4 T cells during tuberculosis
Authors: Sallin Michelle A , Sakai Shunsuke , Kauffman Keith , Young Howard A , Zhu Jinfang (Jeff) , Barber Daniel L 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb) is the leading cause of death by infectious disease worldwide. The development of novel TB vaccination strategies would have dramatic global public health benefits, but have been hindered by an incomplete understanding of the mechanisms of protective immunity against Mtb infection. Recent data indicate that the differentiation state of Th1 cells determines their protective capacity against tuberculosis. We have shown that less-differentiated CXCR3+T-betdim Th1 cells migrate into the lung, expand and control Mtb infection after transfer into infected recipients, while KLRG1+CX3CR1+T-bethigh terminal effectors poorly migrate into the lung, fail to expand and cannot transfer protection against Mtb infection. IL-12/23p40 and T-bet have been shown to promote terminal differentiation, leading to the hypothesis that these host protective factors may have unrecognized detrimental effects on the quality of the Mtb-specific CD4 T cell response. We find IL-12/23p40 promotes Th1 cell expansion and maturation beyond the CD73+CXCR3+T-betdim stage, and T-bet prevents deviation of Th1 cells into Th17 cells. Nevertheless, IL-12/23p40 and T-bet are also essential for the production of a subset of intravascular CX3CR1+KLRG1+Th1 cells. Furthermore, T-bet potently suppresses the development of CD69+CD103+ tissue resident phenotype effectors in the lung. In contrast, CD4 T cell-derived IFNγ inhibits the accumulation of intravascular CX3CR1+KLRG1+Th1 cells. Thus, although IL-12 and T-bet are essential host survival factors, they simultaneously oppose lung CD4 T cell responses at several levels, demonstrating a functional dichotomy of Th1 polarization in tuberculosis.
Alternative polyadenylation signals regulate HLA-A surface expression
Authors: kulkarni smita , Ramsuran Veron , Rucevic Marijana , Lied Alexandra , kulkarni Viraj , Le Gall Sylvie , Carrington Mary 
Published: January 2023

Abstract
Genomic variations in the 5′ and 3′ untranslated region (UTR) have been shown to influence Human leukocyte antigen (HLA) class I expression, the strength of immune responses and disease outcomes beyond peptide binding. Sequencing of the 3′UTR of common HLA-A alleles indicated a presence of three polyadenylation signals (PAS). The proximal and the middle PAS are conserved whereas the distal PAS is disrupted in some alleles. Using 3′ rapid amplification of cDNA ends (3′RACE), we confirmed expression of two distinct forms of HLA-A 3′UTR that use the proximal or the distal PAS and differ in length by 100 base pairs. HLA-A alleles varied in the usage of PAS, with some alleles using only the proximal, and others using both the PAS to differing degrees. The short and long 3′UTRs cloned in luciferase vectors and expressed in the human T cell line-Jurkat showed similar mRNA expression levels. However, the long 3′UTR conferred lower luciferase expression as compared to the short form, indicating translation inhibition of the long 3′UTR. RNA affinity pulldown followed by mass-spectrometric analysis indicated enrichment of an RNA-binding protein (RBP), Syncrip, among proteins bound to the long 3′UTR. Only the long form of cellular HLA-A 3′UTR co-precipitated with Syncrip. The siRNA depletion of Syncrip increased surface expression of HLA-A alleles with the long 3′UTR, whereas an allele expressing only the short form was unaffected. Further, specific blocking of the proximal 3′UTR reduced surface expression without a decrease in mRNA expression. These data demonstrate allele-specific variation in the PAS usage by HLA-A modulated its expression, which adds another layer of regulation and variation to the expression of HLA-A.
JAK/STAT Signaling of Natural Killer (NK) Cells Following Cytokine Stimulation
Authors: Rodriguez-Nieves Jennifer , Tuck Ryan , De Paris Kristina 
Published: January 2023

Abstract
Natural Killer (NK) cells are an important component of the innate immune system, capable of providing a fast and effective response against virally infected cells. NK cells are mainly characterized by their cytotoxic function and their ability to secrete cytokines. It has been shown that infant NK cells have decreased cytotoxicity and cytokine-secreting function, suggesting hyporesponsiveness of NK cells during the first year of life. Because NK cell activation is dependent on cytokine stimulation, we hypothesized that infant NK cells are hyporesponsive due to deficiencies in signaling following cytokine stimulation. We examined the activation of human infant and adult NK cells in response to IL2, IL12 or IFNα stimulation, cytokines important in NK survival and function. Using Phosflow analysis, we measured the phosphorylation of transcription factors, STATs, specific for the distinct cytokines. Cord blood NK cells showed significantly lower pSTAT activation when compared to adult NK cells when treated with IL2 or IFNα but not IL12. However, despite pSTAT4 activation in response to IL12, no nuclear translocation was observed by ImageStreamX Mark II analysis. NK cells in 6 and 12-month-old infants showed similar frequencies of pSTAT NK cells compared to adults when treated with IL2, IL12 or IFNα, suggesting that cytokine signaling in NK cells is age-dependent. Surprisingly, pSTAT activation was restored in cord blood NK cells when treated with a cocktail of IL2, IL12 and IL15. These results suggest that JAK/STAT signaling in infant NK cells is impaired at different steps in the pathway in response to specific cytokines.
Cereblon deficiency increases polyamine synthesis in activated T-cells through the amplification of c-MYC and the arginine and proline metabolic pathway
Authors: Beatty Matthew S , Hesterberg Rebecca S , Han Ying , Epling-Burnette Pearlie 
Published: January 2023

Abstract
Thalidomide, and its immunomodulatory drug analogs (IMiD) including lenalidomide and pomalidomide, activate T-cells and reverse T-cell anergy in cancer patients. Previously we have shown that cereblon, the molecular target of IMiD compounds, is a negative regulator of T-cell metabolism and signaling following suboptimal antigen stimulation. To explore this mechanistically, differentially-regulated genes were subjected to gene set enrichment analysis (GSEA) for transcriptional regulators and global metabolomic profiling was conducted by mass spectrometry in activated Crbn+/+ versus Crbn−/− T-cells. With results confirmed by 1H-NMR spectra analysis, we show that Crbn deficiency leads to an increase in intermediates of the “Arginine/Proline” pathway and enhanced polyamine (putrescine, spermidine, and spermine) biosynthesis. Wang et al showed previously that c-MYC controls polyamine synthesis downstream of mTORC1 during T-cell activation. In activated Crbn−/− CD8+ T-cells, the kinetics and magnitude of c-Myc mRNA were similar to Crbn+/+ T-cells. However, c-MYC is largely regulated through post-transcriptional mechanisms including ubiquitin proteasome-mediated degradation and regulation of c-Myc mRNA translation. Interestingly, polyamines directly bind U- and AU-rich elements (AREs) in the 3′-untranslated region (3′-UTR) of c-Myc mRNA and spermidine is the substrate for a unique post-translational modification required for the efficient translation of c-MYC. Western blot analysis revealed more rapid and sustained expression of c-MYC in activated Crbn−/− T-cells compared to Crbn+/+ T-cells suggesting that CRBN is a regulator of the MYC-polyamine circuit controlling T-cell metabolic fate.
STAT5b Controls IgE-mediated Mast Cell Functions In Vitro and In Vivo
Authors: Kiwanuka Kasalina N , Ryan John J 
Published: January 2023

Abstract
Signal Transducers and Activators of Transcription (STATs) are latent transcription factors that mediate a number of cellular responses including but not limited to cell survival, proliferation and differentiation. This protein family consists of seven members, STATs 1 – 6 including two closely related molecules, STAT5a and STAT5b, that show 96% amino acid sequence homology and are critical for lymphoid, myeloid, and erythroid cell development and function. We recently identified Stat5b as the critical regulator of IgE-mediated cytokine production in mast cells. STAT5b knockout (KO) cells show decreased cytokine production in vitro. In vivo STAT5b KO mice demonstrate decreased sensitivity to IgE-mediated passive systemic anaphylaxis accompanied with decreased production of IL-6 and IL-13 compared to wildtype counterparts. There was no difference in MAR-1-mediated passive systemic anaphylaxis or mast cell number and distribution. Interestingly STAT5b KO mice demonstrate elevated levels of serum IgE and an exacerbated response to histamine-mediated passive systemic anaphylaxis. We previously found that Fyn-mediated Stat5 tyrosine phosphorylation is critical for Stat5-dependent effects. The current work demonstrates that Stat5b is also phosphorylated on serine residues during IgE signaling in mast cells, via a Src-independent pathway requiring PI3-kinase function. Preliminary data show that STAT5b tyrosine and serine phosphorylation either independently or in tandem regulate IgE-induced cytokine production. Elucidating the role of STAT5b in the inflammatory response as well as the key players in STAT5b serine phosphorylation will yield new targets for mast cell-associated diseases.
Establishing the HIV reservoir: HIV-susceptible cells and the signals that recruit them
Authors: Packard Thomas Alexander , Herzig Eytan , Roan Nadia R , Greene Warner C 
Published: January 2023

Abstract
HIV-1 infected individuals require lifelong antiretroviral therapy (ART). If ART is discontinued, HIV rebounds from a reservoir of cells containing latent provirus—persistence of this reservoir is the most significant roadblock to curing people infected with HIV. The HIV reservoir is formed rapidly following infection and remains relatively stable throughout the lifetime of an infected individual on ART: this reservoir persistence likely stems from a central memory phenotype of host T cells.
Transmitted HIV binds and enters cellular targets via the co-receptor, CCR5, which is primarily expressed on effector memory CD4 T cells. How does HIV rapidly establish a central memory reservoir? Abortive HIV infection of lymphoid CD4 T cells activates IFI16, which can form inflammasomes, triggering pyroptosis. IFI16 also can activate STING to produce type 1 interferon and the CCL2 chemokine. Thus, an early innate immune response to HIV sets up a zone of inflammation.
CCL2 attracts cells expressing the receptor CCR2. Strikingly, CCR2 delineates a population of lymphoid CD4 T which co-express CCR5 and markers of central memory (CCR7 & CD62L)—precisely the same type of cells that make up the viral reservoir. We found that these lymphoid CCR2+ T cells migrate in response to CCL2 and are susceptible to infection with R5- or X4-tropic strains of HIV. We propose that CCL2 produced early in HIV infection lures CCR2+ central memory T cells to the site of inflammation. Here these cells can become productively infected and produce virus, or become latently infected and contribute to the stable reservoir. These findings raise the possibility that inflammation contributes to seeding of the latent reservoir, and might be reduced by antagonists of CCL2/CCR2.
The effect of HIV-1 infection on S1P-R1 expression and function in thymocytes and their progenitors during entry into and egress from the human thymus
Authors: Resop Rachel Samantha , Craft Joshua , Vatakis Dimitrios N , Uittenbogaart Christel H 
Published: January 2023

Abstract
Lack of adequate T cell regeneration in HIV infected individuals is likely due to a defect in the entry of hematopoietic stem cells (HSC) into and egress of naïve T cells from the thymus to the periphery. We studied the effect of HIV-1 infection on the receptors to Sphingosine-1-phosphate (S1P), a chemotactic sphingolipid mediator, during thymocyte trafficking. Our novel findings show that HIV infection changes the expression patterns of S1P-R1 on thymocytes about to exit the thymus.
To examine the dynamics of S1P-R1 expression on HSC progenitors and thymocytes during HIV infection, mice implanted with human fetal thymus/liver infected with CXCR4- or CCR5-tropic HIV-1 were injected intravenously with CFSE-labeled HSC. In infected mice CFSE labeled CD34+ progenitors developed into mature thymocytes in the thymic implant and a subset expressed S1P-R1, indicating that entry into the thymus and development are functional during early HIV infection. Surprisingly, we found that S1P-R1 was significantly upregulated in mature thymocytes post-HIV infection. Intriguingly, S1P-R1 was also upregulated by HIV within the CD3+CD69+ population, which normally does not express S1P-R1. S1P-R1 signaling as measured by phosphorylated Akt (pAkt) was not impaired in infected thymocytes, which is interesting in the context of published data demonstrating that S1P-R1 response in HIV infection may be impaired in other cell types.
Further results show that the mechanism of increased S1P-R1 in the thymus by HIV may be due to cytokines including Interferon-beta (IFN-β), which significantly increased S1P-R1 expression in both CD3hiCD69- and CD3+CD69+ thymocyte subsets treated in vitro with exogenous cytokine.
Monitoring the response to TB treatment using T cells biomarkers in TB/HIV patients undergoing ART
Authors: Adekambi Toidi , Ibegbu Chris , Cagle Stephanie , Ray Susan , Rengarajan Jyothi 
Published: January 2023

Abstract
TB is the leading cause of death in HIV-infected persons. Anti-TB treatment in TB/HIV co-infected persons is therefore a public health priority. In patients undergoing anti-TB treatment and ART, ART initiation is often deferred by 2–8 weeks at the physician’s discretion in patients presenting CD4 counts &gt;350 cells/ml to sufficiently reduce the Mtb burden in order to minimize potential drug interactions and complications related to the immune reconstitution inflammatory syndrome (IRIS). However, we currently lack reliable “real time” diagnostic methods to assess Mtb clearance during TB treatment. Isolation of Mtb from sputum can take 4–6 weeks and the rate of sputum smear-negative/active TB cases is upto 66% higher in HIV(+) than in HIV (−) patients. PCR-based technology while sensitive, does not differentiate between live and dead Mtb. We have recently identified specific host immune activation markers (CD38 and HLA-DR) on Mtb-specific CD4+ T cells that correlate with Mtb clearance during and after TB treatment of HIV (−) pulmonary active TB using flow cytometry. We have extended these studies to assess the performance of these biomarkers in diagnosing ATB in HIV(+) patients. Further, we are exploring the value of these biomarker profiles on T cells from TB/HIV co-infected patients undergoing ART and TB treatment, to evaluate their potential to serve as clinical correlates of Mtb clearance in ART treated TB/HIV+ patients.
Germinal Center Immune Dynamics in Secondary Lymphoid Organs after Influenza Vaccination in Children
Authors: Petrovas Constantinos , Amodio Donato , Macchiarulo Giulia , Rocca Salvatore , Cotugno Nicola , Rossi Paolo , Koup Richard , Palma Paolo 
Published: January 2023

Abstract

Introduction
The germinal center immune dynamics leading to protective antibody responses after vaccination in humans remains largely unknown. Investigation of such dynamics would provide valuable information regarding the cellular and molecular mechanisms governing the generation of these responses and lead to novel vaccine strategies

Materials and methods
Paraffin embedded tonsil tissue sections were analyzed by multiplexed confocal microscopy assays and Histo-cytometry. The phenotype of tonsil derived cells was analyzed with polychromatic flow cytometry, flu-specific B cell-responses were detected by ELISPOT to H1N1, H3N2, and B strain and plasmas analyzed with a Luminex multiplex assay.

Results
36 children (24 vaccinated with 2015/2016 seasonal influenza vaccine and 12 controls) scheduled for elective tonsillectomy were included. Flow cytometry and Histo-Cytometry analysis showed that vaccination induced the frequency of tonsil CD4 Tfh, especially the PD-1hiCD57lo ones. This profile was associated with a significant reduction of potential tissue suppressor CD4 T cells and positively correlated with all flu-antigen specific B cells. An induction of IL-4R on plasma cells was found after vaccination implying a further mobilization of these cells. Among the circulating cytokines/chemokines tested, CXCL-13 was selectively induced after vaccination, in line with the increased frequency of CD4 Tfh found.

Conclusions
Considered that children are continuously scheduled for tonsillectomy, tonsils could represent a valuable and approachable secondary lymphoid organ for the investigation of germinal center reactions after vaccination compared to lymph node not easily accessible in this population.
In vitro induction of an atypical memory B cell phenotype in human B cells
Authors: Sohn Hae Won , Ambegaonkar Abhijit A , Obeng-Adjei Nyamekye , Crompton Peter D. , Pierce Susan K 
Published: January 2023

Abstract
Several chronic human infectious diseases including malaria are associated with a large expansion of an unusual population of memory B cells(MBCs) which is referred to as atypical MBCs in malaria. Atypical MBCs in malaria are CD21−, CD27−, CD11c+, FcRL5+, high expression of the transcription factor, T-bet, and have accumulated somatic hyper mutations. These atypical MBCs showed poor B cell receptors signaling and B cell function, ex vivo. These studies suggest that atypical MBCs may contribute to the slow, inefficient acquisition of immunity to malaria in African children. However, the mechanisms by which atypical MBCs are induced during chronic infectious diseases are not known. Recently acute febrile malaria was correlated in African children with increases in peripheral blood TH1-type T follicular helper T cells that secrete inflammatory cytokines, including IFN-γ, and are impaired in their helper function. These results suggested that such T cells or their products may contribute to generation of atypical MBCs. We investigated the conditions under which human B cells can induce an atypical MBC phenotype in vitro. Studying tonsillar B cells we showed that triple combined signaling pathway, BCR, TLR and IFNγ is required for the induction of the malaria-associated atypical MBCs in vitro. In addition, human peripheral blood B cells were induced to express T-bet when incubated anti-Ig and the supernatants of peripheral blood mononuclear cells stimulated by parasite-infected red blood cells. Taken together these results suggest that atypical MBCs may be the product of B cell antigen engagement in the highly inflammatory environments that develop during chronic infections.
IL-17A-producing CD4+Foxp3+ T cells in chronic pulmonary aspergillosis
Authors: Jung Min Kyung , Kang Shin Myung , Kwak Jeong-Eun , Shin Eui-Cheol 
Published: January 2023

Abstract
Chronic pulmonary aspergillosis (CPA) is a severe pulmonary disease caused by aspergillus fumigatus infection. However, immune responses in CPA patients remained to be elucidated. In humans, the CD4+Foxp3+regulatory T cell (Treg) population are classified into the following three subsets based on their expression of CD45RA and Foxp3: CD45RA+Foxp3lo resting Treg cells (subset I), CD45RA−Foxp3high activated Treg cells (subset II), and CD45RA−Foxp3locytokine-secreting non-suppressive cells (subset III). In the present study, we investigated these three CD4+Foxp3+cell subsets and their clinical implication in CPA patients.
The frequency of total CD4+Foxp3+ cells did not differ between CPA patients and healthy controls. Whereas the frequency of subset I or II also did not differ between the two groups, the frequency of subset III was significantly increased in CPA patients compared with healthy controls. Interestingly, IL-17A production was observed in the subset III CD4+Foxp3+ T cells, and the frequency of subset III significantly correlated with IL-17A production from the CD4+ T cell population. Moreover, we found that better clinical prognosis, evaluated by less hospitalization incident, was associated with higher IL-17A production from the CD4+ T cell population and higher frequency of subset III CD4+Foxp3+ cells among CPA patients.
In summary, the frequency of IL-17A-producing CD4+Foxp3+ T cells was increased and correlated with the frequency of IL-17A+ CD4+ T cells, and the higher frequency of IL-17A-producing CD4+Foxp3+ T cells was related with better clinical prognosis. We suggest that IL-17A-producing CD4+Foxp3+ T cells (subset III) may have a role in Th17 regulation and antifungal immune responses in CPA patients.
Sequence and structural analyses reveal distinct and highly diverse human CD8+ TCR repertoires to immunodominant viral antigens
Authors: Chen Guobing , Yang Xinbo , Go Annette , Gao Mingming , Zhang Yongqing , Shi Alvin , Sun Xiaoping , Mariuzza Roy A. , Weng Nan-ping 
Published: January 2023

Abstract
A diverse TCR repertoire is considered as an essential property of T cell immunity to efficiently contain and eliminate viral invasion. However, limited information is available regarding the characteristics of TCR repertoire to a defined viral antigen peptide in human. Here, we report a comprehensive analysis of CD8 TCR repertoires, consensus binding motifs in CDR3, affinities of selected TCR/pMHCs, and crystal structures of two dominant viral antigens (CMV pp65-NLV and IAV M1-GIL) from 8 healthy adults. A few thousands of distinct TCR alpha and beta chains for NLV and GIL were identified containing highly selected public TCRs. The repertoire of NLV-specific TCR was substantially more diverse than that of GIL-specific TCR for both alpha and beta chains. Further analysis of CDR3 amino acid sequences revealed dozens of distinct consensus motifs in alpha and beta chains of both antigen specific TCRs. Single cell analysis showed preference of the CDR3 motifs paring between TCR alpha and beta. The affinity of TCR/pMHC ranged more widely for GIL (Kd, 1.8–191 μM) than for NLV (4.7–28 μM). Structural analysis exhibited similar patterns of three TCR-GIL-HLA-A2 interactions, while two distinct binding patterns among three TCR-NLV-HLA-A2 interactions. Our findings revealed highly diverse viral antigen-specific CD8 TCR repertoire, the consensus binding motifs in CDR3, binding affinity of TCR and pMHC, and different patterns of TCR-pMHC interactions. Such information will be essential to understand the make-up of T cell immunity and to guide the potential immune based clinical intervention such as cell therapy.