bluuebunny/crossref_metadata_2025_split huggingface dataset:
T cells activation and cytokines balance in active and cured tuberculosis (40.7)
Authors: Rodrigues Virmondes , Silva Marcos , Castellano Lucio , Machado Juliana , Figueiredo Amanda , Oliveira Rafael , Faria Gladstone , Pereira Sanivia , Rodrigues Denise 
Published: January 2023

Abstract
Tuberculosis is still a major health problem around the world and the understand of immune response against Mycobacterim tuberculosis is necessary for the development of new strategies for control disease progression. Activation of CD4+ and CD8+ T-cells by flow cytometry and the levels of cytokines in culture supernatants of PBMC from patients with active tuberculosis (n=25) comparing with individuals after successful treatment of tuberculosis (n=22). The activation of CD4+ but not CD8+ T cells was significantly higher in the tuberculosis-treated patients (p=0.04). The levels of IFN-γ and TNF-α were significantly higher among tuberculosis-treated subjects (p=0.005, p=0.034). The levels of cytokines involved in Th-2 and Th-17 axis did not differ among groups. We observed that the levels of IFN-γ and TNF-α were augmented only in subjects after more than 12 months of treatment (p=0.03 and p=0.01). Also, the huge production of Th1 cytokines by cured subjects was followed by an elevation in IL-10 production (p=0.03). Clinical cure due to treatment in tuberculosis was followed by the production of Th1 cytokines, although this recovery does not occur immediately after therapy ending. Th1 cytokine overproduction in healing process is followed by an elevation in IL-10 levels, indicating that concomitantly with elevation of pro-inflammatory cytokines there is a modulation of an immune response regulation-related cytokine.
The role of CD8+ T cells in cutaneous leishmaniasis caused by Leishmania braziliensis (40.9)
Authors: de Oliveira Camila , Novais Fernanda , Santiago Rômulo , Brodskyn Claudia , Barral-Netto Manoel , Barral Aldina 
Published: January 2023

Abstract
The leishmaniases are a group of diseases that affect millions of people worldwide. In the present study, we investigated the participation of CD8+ T cells during infection with L. braziliensis, the main etiolological agent of mucocutaneous leishmaniasis. Following inoculation of L. braziliensis into the ear dermis of BALB/c mice, we observed a high proportion of CD8+CD25+, at the time of lesion appearance, whereas CD8+CD69+ T cells were found at a greater proportion in the draining lymph nodes, at this same time point. Regarding cytokine production, a high percentage of CD8+IFN-g+ cells was detected as early as two weeks post infection, at the ear dermis. The same kinetics was observed for expression of CD107b. Interestingly, presence of IL-4+ and IL-10+ T cells was also detected during the course of infection in the ear dermis. Parasite load in β2-m deficient mice was smaller than in wild type (WT) mice. However, transfer of CD8+ cells obtained from L. braziliensis-primed mice induced the control of lesion development upon transfer to infected mice. These data indicate that CD8+ T cells may play a protective role in cutaneous leishmaniasis caused by L. braziliensis.
Locating the PTPN22 Gene in Families with Multiple Diagnosis of Autoimmune Disorders (41.5)
Authors: Lee Diane 
Published: January 2023

Abstract
While working at NIAID, I examined the PTPN22 gene, which encodes for the lymphoid protein tyrosine phosphatase (PTP), and has been shown to be associated with susceptibility to autoimmune disorders. PTP functions in a coordinated manner with protein tyrosine kinases in the regulation of signaling responses. Kinases are involved with controlling the amplitude of a signaling response and phosphatases are important for controlling the rate and duration of the response. From this laboratory experience, I developed an immunology unit for high school molecular biology and biotechnology classes. This is an intensive month long unit which introduces the first and second lines of defense in the immune system. This unit incorporates routine laboratory practices, such as polymerase chain reaction or PCR, restriction digest and analysis to identify individuals’ susceptibility towards autoimmune diseases. Upon completion of this unit, students will gain an understanding of the immune system through the PTPN22 allele and its effects on autoimmune diseases. The concluding unit activity has students study a family pedigree chart with multiple autoimmune diseases. The PTPN22 gene is isolated from the DNA of non-affected members from this pedigree chart, through the process of PCR. Using restriction digest and analysis, individuals’ DNA is tested to determine the presence of the PTPN22 allele and to establish autoimmune disease susceptibility.
Killer Proteins: Development of a bacteriocidal assay to study the role of complement in killing bacteria. (41.8)
Authors: Long John , Farrar John 
Published: January 2023

Abstract
Complement is an important component in the innate immune system’s defense against bacterial infections. In this unit, we teach basic principles of innate immunity by focusing on the biochemical nature of the complement cascade and its role in directly killing bacteria. We have developed a classroom curriculum that combines didactic course material with a novel laboratory exercise to help students understand the concept of complement in immunology. We have developed a bacteriocidal assay that students will perform to demonstrate the ability of complement to kill bacteria. This laboratory exercise, and subsequent background discussions, will introduce two key aspects of innate immunity. First, complement is a natural group of proteins secreted predominately by the liver which is an effective bacteriocidal agent. Second, complement and antibodies work harmoniously together to recognize and eliminate bacteria as a bridge to adaptive immunity. Students will be able to study the effects of both complement alone and complement plus antibody complexes in this bacteriocidal experiment. An interactive, hands-on laboratory activity enables students to quantify bacterial cell death with a colony forming unit assay of E.coli . Through this exercise, students will be able to focus on basic principles of innate immunity, experimental design and execution, and data analysis.
Increased resistance against MRSA-infection in immunocompromized hosts by injection of IL-12 producing cells. (42.10)
Authors: Saito-Taki Tatsuo , Suka Junko , Onogawa Tsuyoshi , Kitasato Hidero 
Published: January 2023

Abstract
Methicillin-resistant Staphylococcus aureus (MRSA) induces lethal infections at Health-care associated infection in immunocompromized hosts. Since MRSA shows multiple drug resistande, treatments by chemotherapeutic agents are hard to recover from illness. Murine fetal fibroblast cells C57/BL6 (C57/hIL-12) transfected with human IL-12 gene was established and the hIL-12 concentrations in peripheral bloods of hIL-12 were measured in normal beige femal mice and immunodefect beige male mice after ip injection of 1 x 106 of C57/hIL-12 cells and detectable dose of hIL-12 were detected at 12 hr. after injection. Concomitant injections of MRSA and hIL-12 could not induce any protective effects for beige male mice, but preinjection of hIL-12 before 48 hr from MRSA infection induced significant protective effects compared to concomitant injections. These results suggest that 48 hr is critical duration to restore the immune responses of immunodefect beige mice by activating cytokine network via IL-12.
The significance of TLRs in Brucella mediated dendritic cell activation in vitro and pulmonary clearance in vivo (42.16)
Authors: Surendran Naveen , Hiltbold Elizabeth , Sriranganathan Nammalwar , Heid Bettina , Boyle Stephen , Makris Melissa , Zimmerman Kurt , Witonsky Sharon 
Published: January 2023

Abstract
Brucellosis is a worldwide zoonoses affecting 500,000 people annually with no approved human vaccines available. Live attenuated B. abortus vaccine strain RB51 protects cattle through CD4 and CD8 T-cell mediated responses. However, there are concerns regarding its efficacy and use as a live vaccine in people. Therefore, identifying how Brucella vaccines stimulate innate and adaptive immunity is critical to enhancing vaccine efficacy. Brucella stimulate dendritic cells (DCs) through Toll-Like Receptors (TLRs) 2, 4 and 9. In this study, to identify how rough vaccine strains stimulate DC activation and function in vitro, we used vaccine strain RB51 versus pathogenic strain 2308 to stimulate bone marrow derived dendritic cells (BMDCs) from TLR2, 4 or 9 knockout (KO) and wild type BALB/c mice. Since inhalation of infected aerosols is one of the most common routes of exposure, in vivo clearance of strain RB51 compared to strain 2308 from intranasally (IN) infected TLR KO vs. control BALB/c mice was assessed. We determined that strain RB51 induced significant (p≤0.05) DC activation compared to strain 2308 which was TLR independent. However, strain RB51 induced TNF - α production was TLR2 and TLR9 dependent and IL-12 production was TLR2 and TLR4 dependent. TLR4 KO mice had significantly (p≤0.05) delayed pulmonary clearance of strain RB51 in vivo at day 14 post infection. Based on these data, future studies will focus on enhancing the protective ability of strain RB51.
Organization of bovine lambda light chain genes on chromosome 17 (43.14)
Authors: Pasman Yfke , Saini Surrinder , Smith Elspeth , Kaushik Azad 
Published: January 2023

Abstract
Twenty five Vλ genes, seventeen being functional Vλ, corresponding to the three Vλ gene families were identified upon complete characterization and physical mapping of bovine λ-light chain locus on chromosome 17. The bovine λ-light chain locus spans 412 kbp where the Vλgenes are located 23.7 kbp 5’ of four Jλ-Cλ units. The Vλgenes are organized in three subclusters separated by two large introns (126.8 and 138.3 kbp). The predominantly expressed Vλ1 genes are present in the two 5’ sub-clusters, while most Jλ-proximal Vλ sub-cluster comprises rarely expressed Vλ2 and Vλ3 genes. The Jλ3-Cλ3 unit was found predominantly expressed, though Jλ2 and Jλ3 have identical recombination signal sequences (RSS). The Cλ1 and Cλ4 are pseudogenes and their Jλ-Cλ units are not expressed. Endogenous counter selection for Jλ2 encoded CDR3 and framework 4 regions are likely the cause for the predominant expression of Jλ3-Cλ3 genes over Jλ2-Cλ2. The preferential expression of Vλ1 genes in the bovine Immunoglobulin repertoire seems to be influenced by the composition of RSSs. Polymorphism at the λ-light chain locus is responsible for differences in the genomic complexity between Hereford and Holstein cattle. Despite more potential germline encoded combinatorial diversity, the λ-light chain repertoire in cattle is predominantly encoded by Vλ1- Jλ3-Cλ3 recombinations. (Supported by NSERC Canada)
Activation of bovine NK cells through Toll-like receptors (43.2)
Authors: Mayer Sandra , Estes Don , Golde William 
Published: January 2023

Abstract
Toll like receptors (TLRs) play a critical role in the immediate detection and control of infection in mammals and are important for initiating a series of signaling events which result in proinflammatory gene expression. Many of these receptors are type I transmembrane proteins that belong to the IL-1 receptor family. There are 13 TLRs already identified in mammals, 10 in humans, 10 in bovine and 13 in mice. The TLRs main functions include secretion of type I interferon, proinflamatory cytokines, chemokines and expression of co-stimulatory molecules. The aim of this study was to verify intracellular perforin and IFN-γ production, granzyme and perforin transcription and extra cellular IFN-γ secretion in bovine NK cells treated with TLRs ligands such as TLR2, TLR3, TLR4, TLR5 and TLR9. Our preliminary results demonstrated that flagellin (6/6) and PPD (5/6) are the most inducers of intracellular IFN-γ production followed by LPS (4/6) and CpG (4/6). When we analyzed the intracellular perforin production we verified an increase in the cells treated with CpG (4/6) or LPS (3/6). The IFN-γ secretion in the supernatant was analized by a capture ELISA. The Poly I:C had the bigger impact in IFN-γ secretion followed by CpG and LPS which caused a small increase, while PPD and flagellin did not have any effect in IFN-γ secretion. These results suggest that bovine NK cells might be activated by these TLRs ligants, otherwise further studies are needed to elucidate this hypothesis.
Enhancement of Protein Immunogenicity via Synthesis of a Protein-LPS Chimeric Antigen (43.23)
Authors: Ye Jianmin , Kaattari Stephen 
Published: January 2023

Abstract
Typically, while LPS (TI-1) antigens elicit a rapid and robust primary response in teleost fish; they do not provide for good memory. On the other hand protein (TD) antigens, while excellent in priming for memory require much more time for producing a LPS-comparable primary response. This situation is problematic in aquaculture, wherein it is often only feasible to apply a single immunization. To investigate whether a chimeric antigen can produce an accelerated primary response to protein epitopes, while priming for memory, LPS was conjugated to a protein antigen (KLH). The effectiveness of this chimeric antigen (TNP-KLH--LPS) in enhancing an anti-protein response was demonstrated by comparison to protein alone (TNP-KLH) via cellular (elispot) and molecular (titer and affinity) assessments. This enhancement could be due to the increase of ASC number, their increased secretory capacity, or to an altered distribution of ASCs within the body. Furthermore, the chimeric antigen, as opposed to TNP-LPS, appears to elicit a number of higher affinity clones, similar to the protein antigen, which could provide highly specific memory response. Taken together, our results indicate that the chimeric immunogen significantly enhanced protein immunogenicity, which may significantly improve responses to viral subunit vaccines for farmed fish.
Transient Receptor Potential Melastatin 2 (TRPM2) is a Calcium Release Channel Essential for Chemotaxis of Dendritic Cells (44.17)
Authors: Sumoza-Toledo Adriana , Lange Ingo , Bhagat Harivadan , Cortado Hanna , Mori Yasuo , Penner Reinhold , Partida-Sanchez Santiago , Fleig Andrea 
Published: January 2023

Abstract
Chemokines induce Ca2+ signaling and chemotaxis in dendritic cells (DCs) but the molecular players involved in shaping the changes in intracellular Ca2+ remain to be characterized. Using siRNA and knock-out mice, we show that in addition to IP3-mediated Ca2+ release and store-operated Ca2+ entry the transient receptor potential melastatin 2 (TRPM2) channel contributes to Ca2+ release, but not Ca2+ influx in mouse DCs. Consistent with this observation, TRPM2 expression in DCs cells is restricted to endolysosomal vesicles, whereas in neutrophils the channel localizes to the plasma membrane and does not participate in Ca2+ release. While TRPM2 is not involved in DCs maturation, TRPM2-deficient DCs show severely compromised chemotaxis regardless of maturation stage. This is due to perturbed Ca2+ signaling manifested through suppression of specific TRPM2-mediated Ca2+-release events and an additional secondary modification of store-operated Ca2+ entry. DCs deficient in both TRPM2 and inositol-1,4,5-trisphosphate receptor signaling (InsP3R) lose their ability to perform chemotaxis entirely. These findings confirm ADPR as a novel second messenger and highlight TRPM2 as a key player regulating DCs chemotaxis through its function as Ca2+ release channel.
Staphylococcus aureus Biofilm-Mediated Infections: Characterization of the Host Adaptive Immune Response and its Role in Chronic Infection (45.2)
Authors: Prabhakara Ranjani , Harro Janette , Harris Megan , Leid Jeff , Costerton J. , Shirtliff Mark 
Published: January 2023

Abstract
Staphylococcus aureus is a common cause of prosthetic implant infections, which can become chronic due to the ability of S. aureus to grow as a biofilm. Little is known about adaptive immune responses to these infections in vivo. We hypothesized that S. aureus elicits inflammatory Th1/Th17 responses, associated with biofilm formation, instead of protective Th2/Treg responses. A previously developed mouse model of prosthetic implant infection was modified to produce a long term biofilm infection and used to determine chronic infection rates, Treg frequency, and local cytokine levels in B6 and BALB/c mice. Chronic infection levels in STAT6 KO mice (BALB/c background), BALB/c mice receiving anti-CD25 MAb, and B6 mice receiving anti-IL-6 or anti-IL-12 p40 MAb were also assessed. Local S. aureus implant infection was chronic in 100% of B6 mice, whereas BALB/c mice more effectively cleared the infection and demonstrated higher levels of local Th2 cytokines and Tregs. STAT6 KO mice lost the ability to effectively clear an S. aureus implant infection, as did BALB/c mice receiving anti-CD25. In contrast, B6 mice treated with anti-IL-12 p40 were able to more effectively clear the infection. These results indicate that Th2/Treg responses are mechanisms of protection against chronic S. aureus implant infection, as opposed to Th1/Th17 responses, which may play a role in development of chronic implant infection. Our results also suggest that anti-IL-12 p40 MAb may be used therapeutically.
HIV-1-specific exosome targeted CD8+ T cell Vaccine. (45.24)
Authors: Hebbandi Nanjundappa Roopa , Xiang Jim 
Published: January 2023

Abstract
Severe immuno-suppression due to immuno-tolerance and CD4 T cells depletion is the distinguishing feature of HIV-1 infection. Although highly active antiviral therapy (HAART) is available to halt progression of AIDS, it has its own inherent defects, including incomplete protection with associated side effects, and economical concerns. In addition, most of the currently available vaccines fail to induce effective CD8 CTL against HIV-1 in absence of CD4 T cells. Previously, our laboratory showed the efficacy of novel exosome (EXO)-targeted CD4 T cell vaccine (aTexo) with uptake of ova-specific dendritic (DC)-cell-released exosomes in the absence of CD4 T cells using highly metastasizing tumor model. In the present study, we find that aTexo vaccine prepared from active CD8 T cells is capable to induce CD8 CTLs, and its effect is mediated through IL-2 secretion, acquired pMHC I complexes, and partially on CD80 and CD40L costimulators. We have constructed an adenovirus-gp120 vector, which will be used to harvest gp-120-specific exosomes following DC infection, and established various tumor cell lines by transfecting gp-120 gene. We propose to generate gp-120-specific aTexo vaccine and test its efficacy in the presence or absence of CD4 T cells. These studies may significantly impact the development of novel therapeutic vaccines against HIV-1 infection.
Anti-Inflammatory Activity of IVIG Protects Against HSV Encephalitis. (45.26)
Authors: Chandran Ramakrishna , Shen YueWei , Cantin Edouard 
Published: January 2023

Abstract
Susceptible mouse strains like 129 mice develop HSV encephalitis (HSE) characterized by large focal inflammatory CNS lesions associated with massive accumulation of monocytes and neutrophils, resulting in rapid onset of death. In contrast, regulated CNS inflammatory responses protect resistant C57BL/6 mice against HSE. Disease progression was not affected by early inhibition of HSV replication consistent with immune mediated pathology and not virus cytopathology, causing death. Intravenous immunoglobulin (IVIG) possesses potent anti-inflammatory activities and is highly effective in treating various inflammatory diseases. We report here that IVIG protects against fatal HSE by inhibiting CNS inflammation through a mechanism(s) independent of HSV antibody. Both sialylated and non-sialylated but not deglycosylated IgGs mediated protection. IVIG treatment profoundly altered host immune responses, promoting lymphocytic rather than monocytic infiltration of the CNS accompanied by phenotypic alteration of macrophages and a dramatic expansion of CD4+ Tregs that accumulated in peripheral lymphoid organs. However the Tregs were not solely responsible for inhibiting inflammation. IL-10 secreted by APCs was primarily responsible for suppressing CNS inflammation. Despite controlling virus replication IVIG treated IL-10 deficient mice were unable to control CNS inflammation and succumbed eventually. In addition, FcγR2b was critical for IVIG’s immunomodulatory activity.
SIVmac251 infection of Rhesus Macaques destroys secondary lymphoid tissue architecture and depletes naïve T cell populations (45.9)
Authors: Zeng Ming , Wietgrefe Stephen , Schacker Timothy , Southern Peter , Reilly Cavan , Feldman Daniel , Carlis John , Silvestri Guido , Lifson Jeffery , Haase Ashley 
Published: January 2023

Abstract
The organized stromal cell network of secondary lymphoid tissue constitutes a microenvironment that plays a critical role in naïve T cells homeostasis, by providing survival factors such as interleukin-7 (IL-7). We previously showed that collagen deposition and damage to the lymphatic tissue niche in the course of HIV-1 infection contributes to CD4 T cell depletion and limits immune reconstitution with antiretroviral therapy. However, the mechanisms behind it have yet to be defined. Here we show in the SIV-rhesus macaque (Macaca mulatta; RMs) model, that collagen deposition and progressive loss of the stromal cell network limit both access and the source of interleukin-7 on which survival particularly of naïve T cells depends, and, consequently, greatly increased apoptosis results in progressive depletion of both naïve CD4 and CD8 T cell populations. We further show that the loss of the network and T cell depletion are interrelated by the stromal cell network’s dependence on lymphotoxin-beta produced by T cells. Thus, T cell depletion is a multiplicative and progressive pathological process involving access and interdependencies of T cells and supporting lymphatic tissue architecture.
Vaginal herpes simplex virus type 2 infection of mice elicits perforin-dependent immunopathologic tissue damage (46.10)
Authors: Cherpes Thomas , Vicetti Miguel Rodolfo , Kant Jeffrey , Phillips Jaclyn , Schreiner Robert , Harvey Stephen 
Published: January 2023

Abstract
Tissue injury and disease are often caused by host responses to invading pathogens rather than by the pathogen itself. To better understand the mechanisms responsible for immunopathogenic host responses, mice vaginally infected with herpes simplex virus type 2 (HSV-2) were used to model a viral pathogen capable of producing extensive collateral host tissue damage. We observed vaginal HSV-2 infection elicited intense local production of numerous proinflammatory cytokines and chemokines including IFN-gamma, IP-10, and MIP-1 alpha. Beginning 2 dpi, this inflammatory milieu was responsible for recruitment of large numbers of inflammatory monocytes, NK cells, and NKT cells into infected vaginal mucosa. As characterized by their expression of high levels of both perforin and granzyme B, infiltration of these highly activated NK and NKT cells was highly correlated with the onset of genital pathology. Perforin deficient mice cleared primary infection with similar kinetics as wildtype controls while exhibiting significantly decreased levels of genital pathology. The perforin deficient mice were, however, incapable of generating immune responses that prevented lethal dissemination of the virus to the central nervous system. Our data demonstrate a previously unrealized contribution of perforin-mediated mechanisms for HSV-induced tissue damage and may prove useful for exploration of therapeutic interventions that decrease immunopathology without compromising viral clearance.