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Article: Avian Macrophage Responses to Virulent and Avirulent Clostridium perfringens

Kulkarni, Raveendra R. / Gaghan, Carissa / Mohammed, Javid

Pathogens. 2022 Jan. 15, v. 11, no. 1

2022

Abstract: 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+) ... ...

Abstract	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.
Keywords	Clostridium perfringens ; inducible nitric oxide synthase ; interferons ; interleukin-6 ; macrophages ; major histocompatibility complex ; necrotic enteritis ; nitric oxide ; pathogens ; splenocytes ; tonsils ; virulence
Language	English
Dates of publication	2022-0115
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2695572-6
ISSN	2076-0817
ISSN	2076-0817
DOI	10.3390/pathogens11010100
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Mucosal and systemic lymphoid immune responses against

Kulkarni, Raveendra R / Gaghan, Carissa / Gorrell, Kaitlin / Fletcher, Oscar J

Avian pathology : journal of the W.V.P.A

2023 Volume 52, Issue 2, Page(s) 108–118

Abstract: Necrotic enteritis (NE), caused ... ...

Abstract	Necrotic enteritis (NE), caused by
MeSH term(s)	Animals ; Clostridium perfringens/genetics ; Clostridium Infections/veterinary ; Chickens ; Virulence ; Enteritis/veterinary ; Poultry Diseases/pathology ; Immunity ; Anti-Inflammatory Agents/metabolism
Chemical Substances	Anti-Inflammatory Agents
Language	English
Publishing date	2023-01-23
Publishing country	England
Document type	Journal Article
ZDB-ID	1476380-1
ISSN	1465-3338 ; 0307-9457
ISSN (online)	1465-3338
ISSN	0307-9457
DOI	10.1080/03079457.2022.2154195
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Article: Avian Macrophage Responses to Virulent and Avirulent

Kulkarni, Raveendra R / Gaghan, Carissa / Mohammed, Javid

Pathogens (Basel, Switzerland)

2022 Volume 11, Issue 1

Abstract: The present study evaluated the avian macrophage responses ... ...

Abstract	The present study evaluated the avian macrophage responses against
Language	English
Publishing date	2022-01-15
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2695572-6
ISSN	2076-0817
ISSN	2076-0817
DOI	10.3390/pathogens11010100
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Article ; Online: Mucosal and systemic lymphoid immune responses against Clostridium perfringens strains with variable virulence in the production of necrotic enteritis in broiler chickens

Kulkarni, Raveendra R. / Gaghan, Carissa / Gorrell, Kaitlin / Fletcher, Oscar J.

Avian Pathology. 2023 Mar. 04, v. 52, no. 2 p.108-118

2023

Abstract: Necrotic enteritis (NE), caused by Clostridium perfringens, is an economically important disease of chickens. Although NE pathogenesis is moderately well studied, the host immune responses against C. perfringens are poorly understood. The present study ... ...

Abstract	Necrotic enteritis (NE), caused by Clostridium perfringens, is an economically important disease of chickens. Although NE pathogenesis is moderately well studied, the host immune responses against C. perfringens are poorly understood. The present study used an experimental NE model to characterize lymphoid immune responses in the caecal tonsils (CT), bursa of Fabricius, Harderian gland (HG) and spleen tissues of broiler chickens infected with four netB+ C. perfringens strains (CP1, CP5, CP18, and CP26), of which CP18 and CP26 strains also carried the tpeL gene. The gross and histopathological lesions in chickens revealed CP5 to be avirulent, while CP1, CP18, and CP26 strains were virulent with CP26 being “very virulent”. Gene expression analysis showed that, while the virulent strains induced a significantly upregulated expression of pro-inflammatory IL-1β gene in CT, the CP26-infected birds had significantly higher CT transcription of IFNγ and IL-6 pro-inflammatory genes compared to CP5-infected or uninfected chickens. Furthermore, CP26 infection also led to significantly increased bursal and HG expression of the anti-inflammatory/regulatory genes, IL-10 or TGFβ, compared to control, CP5 and CP1 groups. Additionally, the splenic pro- and anti-inflammatory transcriptional changes were observed only in the CP26-infected chickens. An antibody-mediated response, as characterized by increased IL-4 and/or IL-13 transcription and elevated IgM levels in birds infected with virulent strains, particularly in the CP26-infected group compared to uninfected controls, was also evident. Collectively, our findings suggest that lymphoid immune responses during NE in chickens are spatially regulated such that the inflammatory responses against C. perfringens depend on the virulence of the strain.
Keywords	Clostridium perfringens ; birds ; bursa of Fabricius ; cecum ; gene expression ; genes ; histopathology ; interleukin-10 ; interleukin-13 ; interleukin-4 ; interleukin-6 ; models ; necrotic enteritis ; pathogenesis ; spleen ; transcription (genetics) ; virulence ; chickens ; immune response ; cytokines
Language	English
Dates of publication	2023-0304
Size	p. 108-118.
Publishing place	Taylor & Francis
Document type	Article ; Online
ZDB-ID	1476380-1
ISSN	1465-3338 ; 0307-9457
ISSN (online)	1465-3338
ISSN	0307-9457
DOI	10.1080/03079457.2022.2154195
Database	NAL-Catalogue (AGRICOLA)

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Article: Effect of CpG-Oligonucleotide in Enhancing Recombinant Herpes Virus of Turkey-Laryngotracheitis Vaccine-Induced Immune Responses in One-Day-Old Broiler Chickens.

Gaghan, Carissa / Browning, Matthew / Cortes, Aneg L / Gimeno, Isabel M / Kulkarni, Raveendra R

Vaccines

2023 Volume 11, Issue 2

Abstract: Infectious laryngotracheitis (ILT) is an economically important disease of chickens. While the recombinant vaccines can reduce clinical disease severity, the associated drawbacks are poor immunogenicity and delayed onset of immunity. Here, we used CpG- ... ...

Abstract	Infectious laryngotracheitis (ILT) is an economically important disease of chickens. While the recombinant vaccines can reduce clinical disease severity, the associated drawbacks are poor immunogenicity and delayed onset of immunity. Here, we used CpG-oligonucleotides (ODN) as an
Language	English
Publishing date	2023-01-29
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines11020294
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Article: Intracloacal Inoculation of Broiler Chickens with

Gaghan, Carissa / Gorrell, Kaitlin / Taha-Abdelaziz, Khaled / Sharif, Shayan / Kulkarni, Raveendra R

Microorganisms

2023 Volume 11, Issue 3

Abstract: Necrotic enteritis (NE) is an economically important disease of chickens. We have recently shown that inflammatory responses in chickens inoculated orally with ... ...

Abstract	Necrotic enteritis (NE) is an economically important disease of chickens. We have recently shown that inflammatory responses in chickens inoculated orally with virulent
Language	English
Publishing date	2023-03-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms11030771
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Article ; Online: Cellular Immune Responses in Lymphoid Tissues of Broiler Chickens Experimentally Infected with Necrotic Enteritis-Producing

Kulkarni, Raveendra R / Gaghan, Carissa / Mohammed, Javid / Sharif, Shayan / Taha-Abdelaziz, Khaled

Avian diseases

2023 Volume 67, Issue 2, Page(s) 186–196

Abstract: Host cellular responses ... ...

Abstract	Host cellular responses against
MeSH term(s)	Animals ; Clostridium perfringens/physiology ; Clostridium Infections/veterinary ; Clostridium Infections/pathology ; Chickens ; CD8-Positive T-Lymphocytes/pathology ; Enteritis/veterinary ; Enteritis/pathology ; Poultry Diseases/pathology ; Lymphoid Tissue/pathology ; Immunity, Cellular ; Necrosis/veterinary
Language	English
Publishing date	2023-08-09
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	40871-2
ISSN	1938-4351 ; 0005-2086
ISSN (online)	1938-4351
ISSN	0005-2086
DOI	10.1637/aviandiseases-D-23-00012
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Article ; Online: In ovo HVT vaccination enhances cellular responses at hatch and addition of poly I:C offers minimal adjuvant effects.

Boone, Allison C / Kulkarni, Raveendra R / Cortes, Aneg L / Villalobos, Tarsicio / Esandi, Javier / Gimeno, Isabel M

Vaccine

2023 Volume 41, Issue 15, Page(s) 2514–2523

Abstract: In ovo vaccination with herpesvirus of turkey (HVT) hastens immunocompetence in chickens and the recommended dose (RD) of 6080 plaque-forming-units (PFU) offers the most optimal effects. In previous studies conducted in egg-type chickens, in ovo ... ...

Abstract	In ovo vaccination with herpesvirus of turkey (HVT) hastens immunocompetence in chickens and the recommended dose (RD) of 6080 plaque-forming-units (PFU) offers the most optimal effects. In previous studies conducted in egg-type chickens, in ovo vaccination with HVT enhanced lymphoproliferation, wing-web thickness with phytohemagglutinin-L (PHA-L), and increased spleen and lung interferon-gamma(IFN-γ) andToll-like receptor 3 (TLR3) transcripts. Here, we evaluated the cellular mechanisms by which HVT-RD can hasten immunocompetence in one-day-old meat-type chickens, and also determined if HVT adjuvantation with a TLR3 agonist, polyinosinic-polycytidylic acid (poly(I:C)), could enhance vaccine-induced responses and provide dose-sparing effects. Compared to sham-inoculated chickens, HVT-RD significantly increased transcription of splenic TLR3 and IFN γ receptor 2 (R2), and lung IFN γ R2, while the splenic IL-13 transcription was found decreased. Additionally, these birds showed increased wing-web thickness following PHA-L inoculation. The thickness was due to an innate inflammatory cell population, CD3
MeSH term(s)	Animals ; Chickens ; Poly I-C/pharmacology ; Toll-Like Receptor 3 ; Interleukin-13 ; CD28 Antigens ; Marek Disease ; Herpesvirus 1, Meleagrid ; Interferon-gamma ; Vaccination/veterinary
Chemical Substances	Poly I-C (O84C90HH2L) ; Toll-Like Receptor 3 ; Interleukin-13 ; CD28 Antigens ; Interferon-gamma (82115-62-6)
Language	English
Publishing date	2023-03-07
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2023.02.076
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Article ; Online: Immunization of turkeys with Clostridium septicum alpha toxin-based recombinant subunit proteins can confer protection against experimental Clostridial dermatitis.

John, Feba Ann / Criollo, Valeria / Gaghan, Carissa / Armwood, Abigail / Holmes, Jennifer / Thachil, Anil J / Crespo, Rocio / Kulkarni, Raveendra R

PloS one

2024 Volume 19, Issue 4, Page(s) e0302555

Abstract: Clostridial dermatitis (CD), caused by Clostridium septicum, is an emerging disease of increasing economic importance in turkeys. Currently, there are no effective vaccines for CD control. Here, two non-toxic domains of C. septicum alpha toxin, namely ... ...

Abstract	Clostridial dermatitis (CD), caused by Clostridium septicum, is an emerging disease of increasing economic importance in turkeys. Currently, there are no effective vaccines for CD control. Here, two non-toxic domains of C. septicum alpha toxin, namely ntATX-D1 and ntATX-D2, were identified, cloned, and expressed in Escherichia coli as recombinant subunit proteins to investigate their use as potential vaccine candidates. Experimental groups consisted of a Negative control (NCx) that did not receive C. septicum challenge, while the adjuvant-only Positive control (PCx), ntATX-D1 immunization (D1) and ntATX-D2 immunization (D2) groups received C. septicum challenge. Turkeys were immunized subcutaneously with 100 μg of protein at 7, 8 and 9 weeks of age along with an oil-in-water nano-emulsion adjuvant, followed by C. septicum challenge at 11 weeks of age. Results showed that while 46.2% of birds in the PCx group died post-challenge, the rate of mortality in D1- or D2-immunization groups was 13.3%. The gross and histopathological lesions in the skin, muscle and spleen showed that the disease severity was highest in PCx group, while the D2-immunized birds had significantly lower lesion scores when compared to PCx. Gene expression analysis revealed that PCx birds had significantly higher expression of pro-inflammatory cytokine genes in the skin, muscle and spleen than the NCx group, while the D2 group had significantly lower expression of these genes compared to PCx. Peripheral blood cellular analysis showed increased frequencies of activated CD4+ and/or CD8+ cells in the D1 and D2-immunized groups. Additionally, the immunized turkeys developed antigen-specific serum IgY antibodies. Collectively, these findings indicate that ntATX proteins, specifically the ntATX-D2 can be a promising vaccine candidate for protecting turkeys against CD and that the protection mechanisms may include downregulation of C. septicum-induced inflammation and increased CD4+ and CD8+ cellular activation.
MeSH term(s)	Animals ; Turkeys/immunology ; Clostridium septicum/immunology ; Clostridium Infections/prevention & control ; Clostridium Infections/immunology ; Clostridium Infections/veterinary ; Poultry Diseases/prevention & control ; Poultry Diseases/immunology ; Poultry Diseases/microbiology ; Bacterial Toxins/immunology ; Recombinant Proteins/immunology ; Recombinant Proteins/administration & dosage ; Dermatitis/prevention & control ; Dermatitis/immunology ; Dermatitis/veterinary ; Bacterial Vaccines/immunology ; Bacterial Vaccines/administration & dosage ; Immunization
Chemical Substances	Bacterial Toxins ; Recombinant Proteins ; Bacterial Vaccines
Language	English
Publishing date	2024-04-29
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0302555
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Article ; Online: Characterization of immune responses and immunopathology in turkeys experimentally infected with clostridial dermatitis-producing strains of Clostridium septicum.

Criollo, Valeria / John, Feba Ann / Gaghan, Carissa / Fletcher, Oscar J / Thachil, Anil / Crespo, Rocio / Kulkarni, Raveendra R

Veterinary immunology and immunopathology

2024 Volume 269, Page(s) 110717

Abstract: Clostridium septicum is one of the major causative agents of clostridial dermatitis (CD), an emerging disease of turkeys, characterized by sudden deaths and necrotic dermatitis. Despite its economic burden on the poultry industry, the immunopathological ... ...

Abstract	Clostridium septicum is one of the major causative agents of clostridial dermatitis (CD), an emerging disease of turkeys, characterized by sudden deaths and necrotic dermatitis. Despite its economic burden on the poultry industry, the immunopathological changes and pathogen-specific immune responses are poorly characterized. Here, we used three strains of C. septicum, namely Str. A1, Str. B1 and Str. C1, isolated from CD field outbreaks, to experimentally infect turkeys to evaluate local (skin and muscle) and systemic (spleen) pathological and immunological responses. Results showed that while all three strains produced an acute disease, Str. A1 and B1 caused significantly higher mortality when compared to Str. C1. Gross and histopathology evaluation showed that birds infected with Str. A1 and B1 had severe inflammatory, edematous, granulomatous and necrotic lesions in the skin, muscle and spleen, while these lesions produced by Str. C1 were relatively less severe and mostly confined to skin and/or muscle. Immune gene expression in these tissues showed that Str. B1-infected birds had significantly higher expression of interleukin (IL)-1β, IL-6 and interferon (IFN)γ genes compared to uninfected control, suggesting a robust inflammatory response both locally as well as systemically. The transcription of IL-1β and IFNγ in the muscle or spleen of Str. A1-infected birds and IL-1β in the skin of Str. C1-infected group was also significantly higher than control. Additionally, Str. A1 or B1-infected groups also had significantly higher IL-4 transcription in these tissues, while birds infected with all three strains developed C. septicum-specific serum antibodies. Furthermore, splenic cellular immunophenotyping in the infected turkeys showed a marked reduction in CD4+ cells. Collectively, it can be inferred that host responses against C. septicum involve an acute inflammatory response along with antibody production and that the disease severity seem to depend on the strain of C. septicum involved in CD in turkeys.
MeSH term(s)	Humans ; Animals ; Clostridium septicum/physiology ; Clostridium Infections/veterinary ; Turkeys ; Clostridium ; Inflammation/veterinary ; Dermatitis/veterinary ; Immunity ; Poultry Diseases
Language	English
Publishing date	2024-01-24
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	754160-0
ISSN	1873-2534 ; 0165-2427
ISSN (online)	1873-2534
ISSN	0165-2427
DOI	10.1016/j.vetimm.2024.110717
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