@article{boone_kulkarni_cortes_gaghan_mohammed_villalobos_esandi_gimeno_2024, title={Evaluation of Adjuvant Effect of Cytosine-Guanosine-Oligodeoxynucleotide in Meat-Type Chickens Coadministered <i>In Ovo</i> with Herpesvirus of Turkey Vaccine}, volume={2}, ISSN={["1557-8976"]}, url={https://doi.org/10.1089/vim.2023.0125}, DOI={10.1089/vim.2023.0125}, abstractNote={Herpesvirus of turkey (HVT) increases activation of T cells in 1-day-old chickens when administered in ovo. This study evaluated whether adding cytosine-guanosine oligodeoxynucleotides (CpG ODNs) to the HVT vaccine could enhance the adjuvant effect of HVT. We used a CpG ODN dose of 10 μg per egg. The experimental groups were (1) diluent-only control (sham), (2) HVT, (3) HVT+CpG ODN, (4) HVT+non-CpG ODN, (5) CpG ODN, and (6) non-CpG ODN control. Cellular response evaluation included measuring the frequencies of macrophages (KUL01+MHC-II+), gamma delta T cells (γδTCR+MHC-II+), CD4+, and CD8+ T cell subsets, including double-positive (DP) cells. In addition, CD4+ and CD8+ T cell activation was evaluated by measuring the cellular expression of major histocompatibility complex class II (MHC-II), CD44 or CD28 costimulatory molecules. An adjuvant effect was considered when HVT+CpG ODN, but not HVT+non CpG ODN, or CpG ODN, or non-CpG ODN, induced significantly increased effects on any of the immune parameters examined when compared with HVT. The findings showed that (1) HVT vaccination induced significantly higher frequencies of γδ+MHC-II+ and CD4+CD28+ T cells when compared with sham chickens. Frequencies of DP and CD4+CD28+ T cells in HVT-administered birds were significantly higher than those observed in the non-CpG ODN group. (2) Groups receiving HVT+CpG ODN or CpG ODN alone were found to have significantly increased frequencies of activated CD4+ and CD8+ T cells, when compared with HVT. Our results show that CpG ODN administration in ovo with or without HVT significantly increased frequencies of activated CD4+ and CD8+ T cells.}, journal={VIRAL IMMUNOLOGY}, author={Boone, Allison C. and Kulkarni, Raveendra R. and Cortes, Aneg L. and Gaghan, Carissa and Mohammed, Javid and Villalobos, Tarsicio and Esandi, Javier and Gimeno, Isabel M.}, year={2024}, month={Feb} }
 @article{kulkarni_gaghan_mohammed_2022, title={Avian Macrophage Responses to Virulent and Avirulent Clostridium perfringens}, volume={11}, ISSN={["2076-0817"]}, url={https://doi.org/10.3390/pathogens11010100}, DOI={10.3390/pathogens11010100}, abstractNote={The present study evaluated the avian macrophage responses against Clostridium perfringens that varied in their ability to cause necrotic enteritis in chickens. Strains CP5 (avirulent-netB+), CP1 (virulent-netB+), and CP26 (highly virulent-netB+tpeL+) were used to evaluate their effect on macrophages (MQ-NCSU cells) and primary splenic and cecal tonsil mononuclear cells. The bacilli (whole cells) or their secretory products from all three strains induced a significant increase in the macrophage transcription of Toll-like receptor (TLR)21, TLR2, interleukin (IL)-1β, inducible nitric oxide synthase (iNOS), and CD80 genes as well as their nitric oxide (NO) production and major histocompatibility complex (MHC)-II surface expression compared to an unstimulated control. The CP1 and CP26-induced expression of interferon (IFN)γ, IL-6, CD40 genes, MHC-II upregulation, and NO production was significantly higher than that of CP5 and control groups. Furthermore, splenocytes and cecal tonsillocytes stimulated with bacilli or secretory products from all the strains showed a significant increase in the frequency of macrophages, their surface expression of MHC-II and NO production, while CP26-induced responses were significantly higher for the rest of the groups. In summary, macrophage interaction with C. perfringens can lead to cellular activation and, the ability of this pathogen to induce macrophage responses may depend on its level of virulence.}, number={1}, journal={PATHOGENS}, author={Kulkarni, Raveendra R. and Gaghan, Carissa and Mohammed, Javid}, year={2022}, month={Jan} }
 @article{gaghan_adams_mohammed_crespo_livingston_kulkarni_2022, title={Characterization of vaccine-induced immune responses against coccidiosis in broiler chickens}, volume={40}, ISSN={["1873-2518"]}, url={https://doi.org/10.1016/j.vaccine.2022.05.043}, DOI={10.1016/j.vaccine.2022.05.043}, abstractNote={Coccidiosis, caused by Eimeria protozoan species, is an economically important enteric disease of poultry. Although commercial live vaccines are widely used for disease control, the vaccine-induced protective immune mechanisms are poorly characterized. The present study used a commercial broiler vaccine containing a mixture of E. acervulina, E. maxima, and E. tenella. One-day-old chicks were vaccinated by spray followed by a challenge at 21 days of age with a mixture of wild type Eimeria species via oral gavage. Oocyst shedding, immune gene expression and cellular responses in the spleen and cecal tonsils were measured at pre- (days 14 and 21) and post-challenge (days 24, 28 and 35) time points. Results showed that the oocyst counts were significantly reduced in the vaccinated chickens at post-challenge compared to unvaccinated control group. While the vaccinated birds had a significantly increased toll-like receptor (TLR) 21 gene expression at pre-challenge, the transcription of interferon (IFN)γ, Interleukin (IL)-12 and CD40 genes in spleen and cecal tonsils of these birds was significantly higher at post-challenge compared to unvaccinated chickens. Cellular immunophenotyping analysis found that vaccination led to increased frequency of macrophages and activated T cells (CD8+CD44+ and CD4+CD44+) in the spleen and cecal tonsils at post-challenge. Furthermore, in vitro stimulation of chicken macrophages (MQ-NCSU cells) with purified individual species of E. acervulina, E. maxima, and E. tenella showed a significantly increased expression of TLR21, TLR2 and IFNγ genes as well as nitric oxide production. Collectively, these findings suggest that TLR21 and TLR2 may be involved in the immune cell recognition of Eimeria parasites and that the vaccine can induce a robust macrophage activation leading to a T helper-1 dominated protective response at both local and systemic lymphoid tissues.}, number={29}, journal={VACCINE}, publisher={Elsevier BV}, author={Gaghan, Carissa and Adams, Daniel and Mohammed, Javid and Crespo, Rocio and Livingston, Kimberly and Kulkarni, Raveendra R.}, year={2022}, month={Jun}, pages={3893–3902} }
 @article{boyett_crespo_vinueza_gaghan_mohammed_kulkarni_2022, title={Enumeration and speciation of coccidia affecting turkeys using flow cytometry method}, volume={31}, ISSN={["1537-0437"]}, url={https://doi.org/10.1016/j.japr.2022.100270}, DOI={10.1016/j.japr.2022.100270}, abstractNote={Enumeration of Eimeria oocysts is a common practice in monitoring coccidiosis in turkeys; however, the conventional method of manual microscopic examination of Eimeria oocysts is time-consuming. Previously, we used flow cytometry (FCM) to enumerate and speciate coccidia affecting chickens and here, we extended those findings to turkey coccidia species, E. adenoides and E. meleagrimitis. Using FCM, a commercial vaccine containing these species was used to optimize the scatter-plot parameters, including Forward-(size) and Side-(shape/granularity) scatter Area, Height, and Width patterns for Eimeria. The two Eimeria species populations in the vaccine were accurately phenotyped and the gated populations were then sorted using a Cell sorter instrument to obtain pure oocyst suspensions. The individual Eimeria species identity of sorted oocysts was confirmed by PCR using species-specific primers. A significant (P = 0.0466) correlation (R = 0.9893) in the total oocyst count between FCM and manual methods were observed. Furthermore, when FCM was employed to analyze farm fecal samples, the close similarities in the oocyst morphologies coupled with organic debris particulate interference prevented a precise separation of these 2 species resulting in a lack of oocyst count (OPG) correlation between the 2 methods. The OPG counts by FCM were much lower than the manual method; however, a partial OPG trend between the two methods was observed only at the early timepoint collections during a period of 35 d. Collectively, our findings showed that FCM can be used in the enumeration of turkey Eimeria oocysts with a potential scope for a more precise enumeration and speciation in field samples.}, number={3}, journal={JOURNAL OF APPLIED POULTRY RESEARCH}, publisher={Elsevier BV}, author={Boyett, Taylor and Crespo, Rocio and Vinueza, Valeria C. and Gaghan, Carissa and Mohammed, Javid P. and Kulkarni, Raveendra R.}, year={2022}, month={Sep} }
 @article{herrmann_mamo_holmes_mohammed_murphy_bizikova_2022, title={Long-term effects of ciclosporin and oclacitinib on mediators of tolerance, regulatory T-cells, IL-10 and TGF-beta, in dogs with atopic dermatitis}, ISSN={["1365-3164"]}, DOI={10.1111/vde.13140}, abstractNote={AbstractBackgroundAtopic dogs often are managed with allergen‐specific immunotherapy (AIT) and concurrent dosages of ciclosporin (CSA) or oclacitinib to alleviate their clinical signs. Both drugs might affect proper tolerance induction by inhibiting regulatory T‐cell (Treg) induction.Hypothesis/ObjectivesWe evaluated Treg cell numbers and serum interleukin (IL)‐10 and transforming growth factor‐beta (TGF‐β)1 levels in dogs diagnosed with atopic dermatitis (AD) and successfully treated with either CSA or oclacitinib for nine or more months.AnimalsWe included 15 dogs receiving oclacitinib, 14 dogs treated with CSA, 15 healthy dogs, 13 dogs with untreated moderate‐to‐severe AD and 15 atopic dogs controlled with AIT.Materials and MethodsPeripheral blood CD4+CD25+FOXP3+ T‐cell percentages were determined using flow cytometry. Serum concentrations of IL‐10 and TGF‐β1 were measured by enzyme‐linked immunosorbent assay.ResultsThe percentage of Treg cells in the CSA group was significantly lower in comparison with the healthy group (p = 0.0003), the nontreated AD group (p = 0.0056) or the AIT group (p = 0.0186). There was no significant difference in Treg cell percentages between the CSA and oclacitinib groups, nor between the oclacitinib and the healthy, nontreated AD or AIT‐treated dogs. No significant differences were detected in IL‐10 and TGF‐β1 serum concentrations between the five groups.Conclusions and Clinical RelevanceLower Treg cell percentages in the CSA‐treated dogs suggest an impact of this drug on this cell population; however, it does not necessarily mean that it diminishes tolerance. Functionality and cytokine production may be more important than the number of Treg cells. Further studies evaluating the treatment outcome of dogs receiving AIT and concurrent drugs are needed to show clinical relevance.}, journal={VETERINARY DERMATOLOGY}, author={Herrmann, Ina and Mamo, Lisa B. and Holmes, Jenny and Mohammed, Javid P. and Murphy, K. Marcia and Bizikova, Petra}, year={2022}, month={Dec} }
 @article{adams_kulkarni_mohammed_crespo_2022, title={A flow cytometric method for enumeration and speciation of coccidia affecting broiler chickens}, volume={301}, ISSN={["1873-2550"]}, url={https://doi.org/10.1016/j.vetpar.2021.109634}, DOI={10.1016/j.vetpar.2021.109634}, abstractNote={Production losses, mortality, and control measures associated with coccidiosis, caused by Eimera species, cost the broiler industry over $14 billion a year. Current means to distinguish Eimeria species such as oocyst morphology, pre-patent period and site of infection are subjective, labor intensive or unsuitable for high-throughput applications. Although Polymerase Chain Reaction (PCR) techniques have been validated, the target gene cannot differentiate relative abundance of each species in mixed infections. In this study, we developed a non-antibody-based flow cytometry high throughput method to simultaneously enumerate and speciate four Eimeria species, E. acervulina, E. mitis, E. maxima, and E. tenella, using commercial coccidia vaccine as well as field fecal samples. Our findings showed that the four Eimeria oocyst populations could be distinctly speciated based on their size and granularity (shape) via scatter plotting. These distinct populations were sorted and confirmed by quantitative real-time PCR assay. Finally, the flow cytometry findings were applied to enumerate and speciate oocysts from fecal samples collected from commercial broiler flocks vaccinated for coccidiosis at day of hatch and the results were validated against the conventional manual method of floatation and microscopic examination. Collectively, the findings of this study suggested that non-antibody based Flow Cytometry technique can be successful in the simultaneous enumeration and speciation of coccidia. Further development and validation is needed to make this diagnostic tool useful for field applications at a much larger scale as well as to speciate other Eimeria species.}, journal={VETERINARY PARASITOLOGY}, publisher={Elsevier BV}, author={Adams, Daniel S. and Kulkarni, Raveendra R. and Mohammed, Javid P. and Crespo, Rocio}, year={2022}, month={Jan} }
 @article{daneshmand_kermanshahi_mohammed_sekhavati_javadmanesh_ahmadian_alizadeh_razmyar_kulkarni_2022, title={Intestinal changes and immune responses during Clostridium perfringens-induced necrotic enteritis in broiler chickens}, volume={101}, ISSN={["1525-3171"]}, url={https://doi.org/10.1016/j.psj.2021.101652}, DOI={10.1016/j.psj.2021.101652}, abstractNote={Clostridium perfringens-induced necrotic enteritis (NE) is an economically important disease of broiler chickens. The present study evaluated the effect of C. perfringens on the intestinal histomorphometry, enteric microbial colonization, and host immune responses using 3 experimental NE reproduction methods. The experimental groups consisted of 1) unchallenged Control diet (corn-soybean meal), 2) Control diet + Eimera inoculation at d 11 followed by C. perfringens challenge at d 15 (ECp), 3) Wheat-based diet + C. perfringens challenge (WCp), and 4) Wheat-based diet + Eimeria inoculation followed by C. perfringens challenge (WECp). The results showed that chickens receiving ECp and WECp had reduced (P < 0.05) bird performance coupled with enteric gross lesions and epithelial damage at d 17 and 24 of age compared to unchallenged control birds. These ECp and WECp administered birds also had increased (P < 0.05) ileal colonization by clostridia and E. coli at d 17 and 24, while the resident Lactobacillus counts were reduced (P < 0.05) at d 24 of age. Furthermore, at d 24, jejunal transcription of IL-6, IL-10, annexin-A1 and IL-2 genes was upregulated (P < 0.05) in the ECp group, whereas the transcription of TNF receptor associated factor (TRAF)-3 gene was increased (P < 0.05) in WECp treated birds when compared to unchallenged control group. Additionally, stimulation of chicken splenocytes and cecal tonsilocytes with virulent C. perfringens bacilli or their secretory proteins resulted in a higher (P < 0.05) frequency of T cells and their upregulation of MHC-II molecule, as determined by flow cytometry. These findings suggest that C. perfringens, while inducing epithelial damage and changes in microbiota, can also trigger host immune responses. Furthermore, NE reproduction methods using coccidia with or without the wheat-based dietary predisposition seem to facilitate an optimal NE reproduction in broiler chickens and thus, may provide better avenues for future C. perfringens research.}, number={3}, journal={POULTRY SCIENCE}, publisher={Elsevier BV}, author={Daneshmand, Ali and Kermanshahi, Hassan and Mohammed, Javid and Sekhavati, Mohammad Hadi and Javadmanesh, Ali and Ahmadian, Monireh and Alizadeh, Marzieh and Razmyar, Jamshid and Kulkarni, Raveendra R.}, year={2022}, month={Mar} }
 @article{mattner_mohammed_fusakio_giessler_hackstein_opoka_wrage_schey_clark_fraser_et al._2019, title={Genetic and functional data identifying Cd101 as a type 1 diabetes (T1D) susceptibility gene in nonobese diabetic (NOD) mice}, volume={15}, ISSN={["1553-7404"]}, DOI={10.1371/journal.pgen.1008178}, abstractNote={Type 1 diabetes (T1D) is a chronic multi-factorial disorder characterized by the immune-mediated destruction of insulin-producing pancreatic beta cells. Variations at a large number of genes influence susceptibility to spontaneous autoimmune T1D in non-obese diabetic (NOD) mice, one of the most frequently studied animal models for human disease. The genetic analysis of these mice allowed the identification of many insulin-dependent diabetes (Idd) loci and candidate genes, one of them being Cd101. CD101 is a heavily glycosylated transmembrane molecule which exhibits negative-costimulatory functions and promotes regulatory T (Treg) function. It is abundantly expressed on subsets of lymphoid and myeloid cells, particularly within the gastrointestinal tract. We have recently reported that the genotype-dependent expression of CD101 correlates with a decreased susceptibility to T1D in NOD.B6 Idd10 congenic mice compared to parental NOD controls. Here we show that the knockout of CD101 within the introgressed B6-derived Idd10 region increased T1D frequency to that of the NOD strain. This loss of protection from T1D was paralleled by decreased Gr1-expressing myeloid cells and FoxP3+ Tregs and an enhanced accumulation of CD4-positive over CD8-positive T lymphocytes in pancreatic tissues. As compared to CD101+/+ NOD.B6 Idd10 donors, adoptive T cell transfers from CD101−/− NOD.B6 Idd10 mice increased T1D frequency in lymphopenic NOD scid and NOD.B6 Idd10 scid recipients. Increased T1D frequency correlated with a more rapid expansion of the transferred CD101−/− T cells and a lower proportion of recipient Gr1-expressing myeloid cells in the pancreatic lymph nodes. Fewer of the Gr1+ cells in the recipients receiving CD101−/− T cells expressed CD101 and the cells had lower levels of IL-10 and TGF-β mRNA. Thus, our results connect the Cd101 haplotype-dependent protection from T1D to an anti-diabetogenic function of CD101-expressing Tregs and Gr1-positive myeloid cells and confirm the identity of Cd101 as Idd10.}, number={6}, journal={PLOS GENETICS}, author={Mattner, Jochen and Mohammed, Javid P. and Fusakio, Michael E. and Giessler, Claudia and Hackstein, Carl-Philipp and Opoka, Robert and Wrage, Marius and Schey, Regina and Clark, Jan and Fraser, Heather I. and et al.}, year={2019}, month={Jun} }