bluuebunny/crossref_metadata_2025_split huggingface dataset:
A lamprey monoclonal VLR antibody against human CD5 (65.34)
Authors: Yu Cuiling , Ehrhardt Götz , Cooper Max 
Published: January 2023

Abstract
The recently identified VLRB antibodies of the agnathan sea lamprey use highly variable leucine-rich repeats (LRRs) that assume a solenoid shape to bind antigens. Solved structures of monoclonal VLRB antibodies complexed to the H-trisaccharide or HEL show that antigens bind to variable residues in the beta-sheets located on the concave surface of specific VLRB antibodies. In this study we screened clones of a lamprey VLR cDNA expression library prepared from animals immunized with human CD4+ T cells to identify a monoclonal VLRB antibody (T32) that bound to human T lymphocytes, but not mouse T cells. Modulation of the expression levels of the TCR/CD3 complex by mouse antibodies did not affect VLR T32 binding. Staining of T cells, tonsilar B cells and malignant B-CLL cells with VLRB T32 and a mouse anti-CD5 mAb (clone UCHT2) demonstrated a diagonal co-staining pattern for each population of the test cells. Transfection of the CD5-negative OCI-Ly3 cell line with a human CD5 cDNA construct resulted in VLR T32 binding of transfected but not of untransfected cells. Staining of CD5+ cells by VLR T32 was not blocked by pre-incubation with the mouse UCHT2 anti-CD5 mAb, which suggests that they recognizes different CD5 epitopes. We conclude that lamprey monoclonal VLR antibodies offer useful alternatives to conventional antibodies for biomedical purposes.
Agarose slide elisa for diagnosis of HCV-AB in rural areas (65.5)
Authors: Abouelhag Hussien , Safhi Mohammed 
Published: January 2023

Abstract
Hepatitis C (HCV) is a viral infection of the liver affecting 170 million people around the world. Commercially available tests for HCV today are based on enzyme immunosorbent assay (EIA) for detection of HCV-Ab. Regarding the sophisticated equipments required for diagnosis of HCV Thus this work targets offering a dependable field serological test can be used in urban areas as preliminary screening for HCV patients. 150 serum samples were collected from Jazan clinics in Saudi Arabia for HCV patients and 50 serum samples were collected from healthy volunteers. All samples were tested for HCV-Ab using slides coated with 1.5% agarose saturated with HCV Capside synthetic peptide (BIORAD). 50 µl of each sample were applied in spot of Ag saturate agarose and incubated in inverted position in humidified incubator for 90 min at 37\#959;C. After which the slides were washed gently by Tris NaCl buffer PH 7.4 for 20 min. mouse anti human IgG labeled conjugate was added to the slides and incubated in inverted position in humidified incubator for 30 min at 37C. After washing the slides same as before a substrate (BIORAD) was added for 30 min at room temperature then washed out as before. the slides were examined by necked eye and by light microscope against controls. The obtained results were compared with slandered MONOLISA HCV Ag-Ab kite and both results were match. These results proving that agarose slide EIA can be used for preliminary diagnosis of HCV in field clinics of rural areas.
Near-infrared in vivo imaging of human lymphatic architecture and function in lymphedema and normal subjects (65.7)
Authors: Aldrich Melissa , Adams Kristen , Angermiller Bonnie , Darne Chinmay , Fife Caroline , Guilliod Renie , Hawkins Trina , Marshall Milton , Maus Erik , Rasmussen John , Smith Latisha , Tan I-Chih , Wang XueJuan , Sevick-Muraca Eva 
Published: January 2023

Abstract
Lymphedema (LE) is a disabling condition resulting from genetic predisposition, cancer treatments involving lymph nodes, or nematode infection. Obtaining in vivo images of healthy and diseased lymphatic vessels and lymph nodes, until now, has required using radionucleotide imaging agents, and has resulted in grainy, nondistinct images. We used a custom-designed ICCD camera system and low-level laser excitation of intradermally injected fluorescent dye to obtain live, non-invasive images of lymphatic vessel architecture and function in humans with and without lymphedema. This technique has provided exquisite images and movies of lymphatic abnormalities such as tortuous vessels and patchy “starry night” areas, as well as the ability to measure lymph propulsive velocity and frequency. This non-invasive, live, in vivo imaging of the human lymphatic system in health and disease may provide opportunities to diagnose lymphatic disorders, direct cancer resections, and evaluate therapies.
Dendritic cells support long term survival of plasma cells via CD28-CD80/CD86 interactions (66.11)
Authors: Nair Jayakumar , Carlson Louise , Rozanski Cheryl , Koorella Chandana , Murray Megan , Shaughnessy John Jr , Bergsagel P Leif , Boise Lawrence , Chanan-Khan Asher , Lee Kelvin 
Published: January 2023

Abstract
We have previously shown (AAI2009) that dendritic cells (DC) contribute to the survival of long lived plasma cells in the bone marrow, which are important to maintain persistent antibody titres. What is not clear are the molecular interactions that are involved. Both normal and malignant plasma cells(like myeloma cells) express the T-cell co-stimulatory molecule CD28, and both interact with DCs in the bone marrow microenvironment. CD28 activation on MM triggers similar molecular partners downstream as observed in T-cells. Gene expression analysis of normal and malignant plasma cells shows that CD28 expression correlates with poor prognostic subgroups that are associated with specific genetic lesions. In our data, while activation of CD28 with antibodies or DC-CD80/CD86 (B7) protects myeloma cells against cell death, the ligation of B7 on DCs induces DC production of plasma cell survival factor IL-6, and immunosuppressive factor indoleamine 2,3-dioxygenase (IDO). Use of Lenalidomide (a common myeloma drug) to inhibit DC-B7 expression or anti-CD28(Fab)2 fragments to block CD28-B7 interactions reduced DC ability to protect MM cells. Experiments with αCD28(Fab)2 or inhibitors of notch pathway suggest a cross talk that regulates IL-6 production. IDO activity in DC-MM cocultures, suppressed T-cell proliferation in MLRs while an IDO inhibitor alleviated this. Our data suggests a role for CD28-B7 in plasma cell survival by modulating the bone marrow microenvironment.
A small molecule TGF-b signaling inhibitor synergizes with agonistic OX40 antibody to elicit a potent anti-tumor and anti-metastatic response in a breast cancer model (66.22)
Authors: Garrison Kendra , Zhang Xiamei , Lee Wen-Cherng , Ling Leona , Akporiaye Emmanuel 
Published: January 2023

Abstract
SM16 is an orally available small molecule TGF-b signaling inhibitor that binds to the ATP pocket of TGF-b receptor Type 1 and prevents phosphorylation by TGF-BRII following engagement of the ligand. OX40 (CD134) is a member of the TNF-a receptor superfamily and is upregulated on activated CD4 and CD8 T cells. OX40 ligation by its cognate ligand (OX40L) or agonistic antibody (anti-OX40) enhances effector function, expansion and survival of activated T cells. In this study, we examined the therapeutic efficacy and anti-tumor immune response induced by the combination of SM16 plus anti-OX40 in a poorly immunogenic and highly metastatic 4T1 mammary tumor model. Our data show that SM16 synergizes with OX40 to elicit a potent anti-tumor effect against established primary tumors, with a 73% reduction in tumor size and a 92% reduction in the number of metastatic lung nodules; overall 71% of treated mice were either tumor free or showed stable disease by the end of the study. This positive treatment outcome is associated with a two-fold expansion of tumor-infiltrating effector CD8+ T cells and an accumulation of CD4+ and CD8+ cells overall, as well as a high tumor-specific IFN-g response. Depletion of both CD4+ and CD8+ T cells completely abrogates the effect of this therapy, indicating it is T cell dependent. Taken together, these data suggest that combining a TGF-b signaling inhibitor with anti-OX40 is a viable approach for treating metastatic breast cancer.
Myeloid derived suppressor cells induced thymic iNKT17 cells that promote tumor metastasis (66.36)
Authors: Liu Yuelong , Cheng Ziqiang , Wang Guijun , Sun Dongmei , Liu Cunren , Zhuang Xiaoying , Zhang Huang-Ge 
Published: January 2023

Abstract
A new subset of NKT cells, an IL-17-producing iNKT cell (iNKT17), has been identified recently. The role of this new iNKT17 subset in a tumor bearing host is unknown. We report that the percentages of iNKT17 cells in the thymus are increased dramatically in naïve mice from 0.5 ± 0.1% to 2.1 ± 0.2% in 4T1 breast tumor-bearing mice. Tumor induced iNKT17 cells are marked as TCRβint CD1dTetramerpos CD4neg CD8neg CD44hi CD25hi CD5pos CD24low DX5neg CD54pos. The iNKT17 cells from 4T1 tumor bearing mice are unique in that upon α-Gelcer-CD1d tetramer stimulation, IL-17 but not IL-4, IFN-γ, IL-10, IL-13, IL-6, IL-12 is induced. The CD11b+Gr1+ myeloid-derived suppressor cells (MDSC) accumulate in dramatic fashion in the thymus in 4T1 tumor bearing mice. The induction of iNKT17 subset is MDSC dependent. Addition of antibodies against TGFβ or IL-6 leads to the inhibition of MDSC-mediated induction of iNKT17 cells, suggesting that TGFβ and IL-6 are required for the induction. The MDSC-mediated induction of the iNKT17 cell subset is correlated with reduction of SLAM but not Ly9, CD84, Ly108 and 2B4. Mice injected intravenously with anti-IL17 antibody leads to the attenuation of 4T1 tumor metastasis to lung. This data suggest that MDSC-mediated induction of iNKT17 cells may play a role in promoting tumor metastasis.
Antibody responses to dengue infection (67.9)
Authors: Dejnirattisai Wanwisa , Malasit Prida , Mongkolsapaya Juthathip , Screaton Gavin 
Published: January 2023

Abstract
Dengue virus infection is a major public health problem in tropical and subtropical countries. Around 50 million people are infected per year of which 500,000 cases develop into the severe form, dengue hemorrhagic fever (DHF). Dengue is a flavivirus circulating as 4 serotypes with an average of 30% amino acid difference between each other, and infection with one serotype dose not provide protective immunity to the others, so secondary infection is very common. So far, the mechanisms involved in the pathogenesis of DHF are not well understood, but the key epidemiological studies indicate that it usually occurs in subsequent heterotypic dengue infection. Antibody dependent enhancement (ADE) is one of the hypotheses where preexisting antibodies from primary infection may be unable to neutralise other serotype from secondary infection but may instead form dengue/antibody complexes which will be taken up by Fc-bearing cells such as monocytes and macrophages leading to increase viral production. In this study, we have generated a panel of human monoclonal antibodies against dengue virus from B cells isolated from dengue infected patients. Neutralizing, enhancing and cross-reactive activity of these antibodies have been characterized and shown different patterns of functioanl properties.
Plasmodium chabaudi infection alters B cell homeostasis and disrupts development of heterologous immunity (99.18)
Authors: Wilmore Joel , Maue Alexander , Rochford Rosemary 
Published: January 2023

Abstract
Several studies have shown that the antibody responses to several vaccines are reduced in children following an acute episode of Plasmodium malaria. At present, the factors underlying these reduced responses are not understood. Using the Plasmodium chabaudi mouse model of blood stage malaria, we show that there are dramatic alterations in B cell hematopoiesis in the spleen and bone marrow. Production of immature B cells in the bone marrow was significantly reduced compared to control mice at 12 and 17 days post-infection resulting in significant reductions in transitional, marginal zone and an overall decrease of splenic B cells. To determine if these changes in B cell hematopoiesis impact the development of a heterologous immune response, we immunized P. chabaudi-infected mice during the acute phase of parasitemia with the model vaccine antigen, NP-ovalbumin. At 12 days post immunization, antigen-specific antibody titers of infected mice were significantly lower compared to uninfected immunized mice. Similar numbers of NP-OVA binding B cells could be isolated from infected and control mice. However, P. chabaudi infected mice had significantly reduced levels of BrdU uptake in NP-OVA+ B cells indicating an impaired ability of NP-OVA specific B cells to proliferate. These data suggest that disruption of B cell homeostasis during acute P. chabaudi infection impacts the capacity of the immune system to respond to heterologous infection and immunization.
Variability in murine airway immune responses to clinical and environmental isolates of Aspergillus conidia (99.21)
Authors: Templeton Steven , Buskirk Amanda , Law Brandon , Green Brett , Beezhold Donald 
Published: January 2023

Abstract
Pulmonary exposure to conidia of Aspergillus species has been associated with morbidity and mortality. However, the potential to cause invasive disease varies between species. Results from a previous study in our laboratory suggest that murine airway immune responses are species-dependent, as airway recruitment of CD8+ T effector and memory-phenotype cells were increased in response to aspiration of viable A. fumigatus, but not A. versicolor, conidia. We sought to test the hypothesis that differences in airway responses are strain dependent, by testing additional clinical and environmental Aspergillus isolates in our model. Airway CD4+ or CD8+ T cells produced IFN-γ and/or IL-17A in response to both clinical and environmental strains of A. fumigatus. In A. versicolor aspirated mice, the T-cell response was characterized by lower numbers of both cytokine-producing cells and infiltrating CD8+ T cells. Both A. fumigatus and A. versicolor induced bronchiocentric inflammation and granuloma formation observed in histopathology. The ability of aspirated conidia to germinate in the lung tissue was variable. Airway eosinophil recruitment was increased in response to A. versicolor and a single environmental isolate of A. fumigatus in comparison with other A. fumigatus isolates. These data provide preliminary evidence that airway immune responses to Aspergillus conidia are both species- and strain-dependent.
CCR6+Tc17 cells induced in the absence of CD4 T-cell help upon vaccination with Blastomyces dermatitidis mediate protective immunity (99.8)
Authors: Nanjappa Som , Heninger Erika , Wuthrich Marcel , Klein Bruce 
Published: January 2023

Abstract
The increasing incidence of fungal infections in immune-compromised patients calls for the development of suitable vaccines to target CD8 T cells. Accumulating studies indicate that Th17 cells play an important role in anti-fungal immunity. However, whether IL-17 producing CD8 T cells (Tc17) can be induced and mediate protective immunity against fungal infection in the absence of CD4 help is not known. Here, using mouse model of vaccination against lethal Blastomyces dermatitidis infection, we show that upon vaccination Tc17 cells are induced and re-called at the site of lethal challenge in the absence of CD4 T-cell help. Vaccine immunity by CD8 T cells was severely impaired in both vaccinated- IL-17A KO and IL-17-receptor KO mice in the absence of CD4 T cells. Neutralization of IL-17A during the re-call response also impaired fungal clearance by effector CD8 T cells. Further, we show that protective immunity by Tc17 cells was not influenced by deficient IL-12 signaling, required for Type I immunity, but neutrophils were essential in Tc17 cell action. Tc17 cells preferentially expressed CCR6 over CXCR3 chemokine receptor (distinct from IFNγ producing cells, which were CXCR3+ve) and antibody blocking of CXCR3 did not affect the re-call response of Tc17 cells to the lung. Collectively, our studies show that protective Tc17 cells are induced in the absence of CD4 T cell help and may aid in designing fungal vaccines for immune-compromised hosts.
Type I IFN drives a distinctive dendritic cell maturation phenotype that allows continued MHC-II synthesis and antigen processing (106.21)
Authors: Wearsch Pamela , Simmons Daimon , Canaday David , Liu Yi , Wang Ying , Boom W. , Harding Clifford 
Published: January 2023

Abstract
Microbial molecules or cytokines can stimulate dendritic cell (DC) maturation, which involves DC migration to lymph nodes and enhanced presentation of Ag to launch T cell responses. Toll-like receptor (TLR) agonists are the most studied inducers of DC maturation and promote a burst of Ag processing with enhanced expression of peptide-MHC-II complexes and co-stimulatory molecules. Subsequently, MHC-II localization is shifted from vacuolar compartments to the surface and synthesis is inhibited, thus decreasing Ag processing and limiting presentation to a cohort of Ags kinetically associated with the maturation signal. Type I interferon (IFN-I) also promotes DC maturation, but the phenotypic changes induced by IFN-I are poorly defined. Our studies show that treatment of DC with IFN-I also increases MHC-II Ag presentation, but, in contrast to TLR-induced maturation, allows for continued Ag processing. IFN-I treated DC expressed high levels of MHC-II at the surface as well as in vacuolar compartments, consistent with sustained Ag processing function. Expression of mRNA for MHC-II and CIITA was inhibited in DC treated with TLR agonists, but maintained in DC matured with IFN-I. Similar to TLR agonists, however, IFN-I induced changes in MARCH1 expression and MHC-II ubiquitination. Our findings suggest that IFN-I drives a distinctive DC maturation program that enhances Ag presentation to T cells without a shutdown of Ag processing, allowing continued sampling of Ags for presentation.
Components of the cytoplasmic antigen processing pathway regulates indirect antigen presentation by MHC class II molecules (106.23)
Authors: Joyce Sebastian , Dragovic Srdjan , Hill Timothy , Christianson Gregory , Roopenian Derry , Van Kaer Luc 
Published: January 2023

Abstract
Indirect presentation is a process in which MHC class II-positive antigen-presenting cells (APCs) acquire and present antigens expressed by class II-negative allografts, tumours or infected cells. It is generally assumed that similar cellular biochemical mechanisms underlie direct and indirect antigen presentation. This is because in both cases antigens are delivered to the endo/lysosomes for proteolytic processing for assembly of derived peptides with class II molecules and their subsequent display on the cell surface of antigen presenting cells. Remarkably, upon testing this assumption, we found that indirect presentation involved LMP2-dependent immunoproteasomes, indicating that the antigen had accessed the recipient APC’s cytosol. In this cytosol, TAP and ERAAP, but not tapasin, partially destroyed the antigens slated for indirect presentation. Because autophagy was not involved in returning the donated antigen from the cytosol to the endo/lysosome, a novel mechanism(s) is predicted to underlie cytosol to endo/lysosome antigen transport. How components of the cytosolic antigen-processing pathway regulate indirect class II-restricted antigen presentation and the mechanism(s) by which cytosolic antigens are returned to the endo/lysosomes will be presented.
Classical antigen presentation by HLA-A2 of Mycobacterium tuberculosis derived ligands (106.40)
Authors: Liles Lauren , McMurtrey Curtis , Schafer Fredda , Bardet Wilfried , Swarbrick Gwendolyn , Null Megan , Toren Katelynne , Lewinsohn David , Lewinsohn Deborah , Hildebrand William 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb) remains a world health threat, with 8.8 million cases reported in 2010, 65% of which were new and recurrent cases of tuberculosis . Cellular immunity is critical for controlling Mtb and recent studies identify CD8+ T cells as important players in controlling infection. Cytotoxic T lymphocytes (CTLs) recognize peptide antigen presented by Major Histocompatibility Complex (MHC) Class I molecules and destroy infected cells, therefore discovery of CTL epitopes is of high importance. Using a method of soluble HLA production, we were able to generate soluble HLA-A2 from THP-1 monocyte cell lines transfected with a soluble form of HLA A*02:01. Transfected cells were seeded into a hollow fiber bioreactor and were infected with Mtb or left uninfected. Following production of 25 mgs of HLA-A2 from infected and uninfected cells, peptides were eluted from the supernatants of uninfected and infected cells. Peptides from each group were fractionated using HPLC and analyzed by mass spectrometry. We were able to identify six Mtb derived ligands presented by the MHC Class I molecule HLA-A*0201, two of which reacted well with donor PBMC from patients with both active and latent Mtb infection. Identifying new vaccine candidates is a top priority in the TB field and the reactive Mtb ligands identified here demonstrate that multiple Mtb proteins are processed for presentation by Class I HLA and a fraction of these ligands are immunologically reactive.
Immunization strategies for high affinity immune response (106.48)
Authors: Hawksworth David , Bax Brian , Himmelsbach Richard 
Published: January 2023

Abstract
In the field of hybridoma development immunization strategies involving varied mouse strains, adjuvants and immunogens have been widely reported with diverse results. We describe here studies completed to evaluate two arms of the immunization strategy - adjuvant selection and hapten-to-carrier ratio. First, Titanium Powder Adjuvant (TPA) and 2-Dose Adjuvant (2DA) were evaluated vs. our standard adjuvant, Adjulite Freund’s Adjuvant (AFA), for resulting antibody titer and affinity by immunizing with either insulin or hepatitis B surface antigen (HBsAg). Second, mice were immunized with the hapten neopterin linked to BSA using a 23:1 or 4:1 coupling ratio. CAF1/J and RBF/DnJ mice were employed for both studies. Adjuvant study results showed a higher titer and higher affinity response using AFA vs. TPA. While all mice immunized with HBsAg developed an immune response, those immunized using AFA showed a significantly higher average relative affinity than those immunized using TPA. Additionally, for CAF1/J mice immunized with insulin, only those using AFA developed an immune response. The 2DA mice yielded a relative affinity similar to the AFA mice. In the hapten-to-carrier ratio study mice immunized with hapten coupled using a 4:1 ratio yielded a significantly higher average affinity. We continue to evaluate all arms of the immune response strategy to ensure a high affinity immune response by testing new adjuvants and animal models.