Specific antibody responses against Neospora caninum recombinant rNcGRA7, rNcSAG4, rNcBSR4 and rNcSRS9 proteins are correlated with virulence in mice

SUMMARYThe intraspecific diversity of Neospora caninum is a determinant for in vivo parasite virulence and in vitro parasite behaviour. The relationship between isolate virulence and specific antibody responses against key parasite proteins has not been well characterized. The response kinetics and the differences in specific anti-rNcGRA7, -rNcSAG4, -rNcBSR4 and -rNcSRS9 antibody levels were analysed by recombinant protein-based ELISA in groups of mice inoculated with 10 different N. caninum isolates that differ in their virulence. The majority of the virulence parameters analysed correlated with the specific antibody levels against the 4 recombinant proteins. The antibodies developed against the highly immunogenic protein NcGRA7 were significantly higher in mice inoculated with high virulence isolates than in those inoculated with low-to-moderate virulence isolates in both non-pregnant and pregnant mouse models. Moreover, these levels were correlated with the anti-N. caninum IgG1 and IgG2a responses and the in vitro tachyzoite yield at 56 h. The antibodies directed against the bradyzoite-specific proteins were not detected in a non-pregnant mouse model. However, some seropositive mice were found in groups inoculated with high virulence isolates in a pregnant mouse model. NcGRA7 and NcSAG4 are proteins clearly correlated with virulence, and to a lesser extent NcBSR4 and NcSRS9 proteins. Moreover, antibodies to bradyzoite-specific proteins appear to also be related to virulence in mice. Further analyses should be performed in order to verify the usefulness of these proteins as predictive markers for virulence in an experimental bovine model of neosporosis.

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Characterization of an Outbred Pregnant Mouse Model of Neospora caninum Infection

Journal of Parasitology ◽

10.2307/3285344 ◽

2002 ◽

Vol 88 (4) ◽

pp. 691

Author(s):

Helen E. Quinn ◽

Catherine M. D. Miller ◽

Cheryl Ryce ◽

Peter A. Windsor ◽

John T. Ellis

Keyword(s):

Mouse Model ◽

Neospora Caninum ◽

Pregnant Mouse ◽

Caninum Infection

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Enhancement of in vitro and in vivo antigen-specific antibody responses by interleukin 11.

Journal of Experimental Medicine ◽

10.1084/jem.175.1.211 ◽

1992 ◽

Vol 175 (1) ◽

pp. 211-216 ◽

Cited By ~ 75

Author(s):

T G Yin ◽

P Schendel ◽

Y C Yang

Keyword(s):

T Cells ◽

Antibody Titer ◽

Specific Antibody ◽

Antibody Responses ◽

Rabbit Serum ◽

Dependent Manner ◽

Dose Dependent ◽

Interleukin 11

The availability of large quantities of highly purified recombinant interleukin 11 (rhuIL-11) has allowed us to investigate the effects of rhuIL-11 on sheep red blood cell (SRBC)-specific antibody responses in the murine system. The results showed that rhuIL-11 was effective in enhancing the generation of mouse spleen SRBC-specific plaque-forming cells (PFC) in the in vitro cell culture system in a dose-dependent manner. These effects of rhuIL-11 were abrogated completely by the addition of anti-rhuIL-11 antibody, but not by the addition of preimmunized rabbit serum. Cell-depletion studies revealed that L3T4 (CD4)+ T cells, but not Lyt-2 (CD8)+ T cells, are required in the rhuIL-11-stimulated augmentation of SRBC-specific antibody responses. The effects of rhuIL-11 on the SRBC-specific antibody responses in vivo were also examined. RhuIL-11 administration to normal C3H/HeJ mice resulted in a dose-dependent increase in the number of spleen SRBC-specific PFC as well as serum SRBC-specific antibody titer in both the primary and secondary immune responses. In mice immunosuppressed by cyclophosphamide treatment, rhuIL-11 administration significantly augmented the number of spleen SRBC-specific PFC as well as serum SRBC-specific antibody titer when compared with the cyclophosphamide-treated mice without IL-11 treatment. These results demonstrated that IL-11 is a novel cytokine involved in modulating antigen-specific antibody responses in vitro as well as in vivo.

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Monoclonal anti-interleukin-6 receptor antibody attenuates donor-specific antibody responses in a mouse model of allosensitization

Transplant Immunology ◽

10.1016/j.trim.2013.03.003 ◽

2013 ◽

Vol 28 (2-3) ◽

pp. 138-143 ◽

Cited By ~ 19

Author(s):

G. Wu ◽

N. Chai ◽

Irene Kim ◽

A.S. Klein ◽

S.C. Jordan

Keyword(s):

Mouse Model ◽

Specific Antibody ◽

Interleukin 6 ◽

Antibody Responses ◽

Receptor Antibody ◽

Interleukin 6 Receptor

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CHARACTERIZATION OF AN OUTBRED PREGNANT MOUSE MODEL OF NEOSPORA CANINUM INFECTION

Journal of Parasitology ◽

10.1645/0022-3395(2002)088[0691:coaopm]2.0.co;2 ◽

2002 ◽

Vol 88 (4) ◽

pp. 691-696 ◽

Cited By ~ 28

Author(s):

Helen E. Quinn ◽

Catherine M. D. Miller ◽

Cheryl Ryce ◽

Peter A. Windsor ◽

John T. Ellis

Keyword(s):

Mouse Model ◽

Neospora Caninum ◽

Pregnant Mouse ◽

Caninum Infection

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Expansion of SARS-CoV-2-specific Antibody-secreting Cells and Generation of Neutralizing Antibodies in Hospitalized COVID-19 Patients

10.1101/2020.05.28.118729 ◽

2020 ◽

Cited By ~ 1

Author(s):

Renata Varnaitė ◽

Marina García ◽

Hedvig Glans ◽

Kimia T. Maleki ◽

John Tyler Sandberg ◽

...

Keyword(s):

B Cell ◽

Specific Antibody ◽

Neutralizing Antibodies ◽

Cell Activation ◽

Neutralizing Antibody ◽

Antibody Responses ◽

B Cell Activation ◽

Immunological Parameters ◽

Antibody Secreting Cell ◽

Antibody Levels

AbstractCoronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), emerged in late 2019 and has since become a global pandemic. Pathogen-specific antibodies are typically a major predictor of protective immunity, yet B cell and antibody responses during COVID-19 are not fully understood. Here, we analyzed antibody-secreting cell (ASC) and antibody responses in twenty hospitalized COVID-19 patients. The patients exhibited typical symptoms of COVID-19, and presented with reduced lymphocyte numbers and increased T cell and B cell activation. Importantly, we detected an expansion of SARS-CoV-2 nucleocapsid protein-specific ASCs in all twenty COVID-19 patients using a multicolor FluoroSpot assay. Out of the 20 patients, 16 had developed SARS-CoV-2-neutralizing antibodies by the time of inclusion in the study. SARS-CoV-2-specific IgA, IgG and IgM antibody levels positively correlated with SARS-CoV-2-neutralizing antibody titers, suggesting that SARS-CoV-2-specific antibody levels may reflect the titers of neutralizing antibodies in COVID-19 patients during the acute phase of infection. Lastly, we showed that interleukin 6 (IL-6) and C-reactive protein (CRP) concentrations were higher in serum of patients who were hospitalized for longer, supporting the recent observations that IL-6 and CRP could be used to predict COVID-19 severity. Altogether, this study constitutes a detailed description of clinical and immunological parameters in twenty COVID-19 patients, with a focus on B cell and antibody responses, and provides tools to study immune responses to SARS-CoV-2 infection and vaccination.

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Genetic manipulation of Neospora caninum to express the bradyzoite-specific protein NcSAG4 in tachyzoites

Parasitology ◽

10.1017/s0031182010001666 ◽

2011 ◽

Vol 138 (4) ◽

pp. 472-480 ◽

Cited By ~ 9

Author(s):

V. MARUGÁN-HERNÁNDEZ ◽

L. M. ORTEGA-MORA ◽

A. AGUADO-MARTÍNEZ ◽

G. ÁLVAREZ-GARCÍA

Keyword(s):

Mouse Model ◽

Genetic Manipulation ◽

Neospora Caninum ◽

Constitutive Expression ◽

Chronic Phase ◽

Specific Protein ◽

Host Cells ◽

Mouse Model Of Infection ◽

Transgenic Strains

SUMMARYNeospora caninum is an apicomplexan parasite and the aetiological agent of bovine neosporosis, one of the main causes of reproductive failure worldwide. We have generated 2 independent transgenic knock-in clones, Nc-1SAG4c1.1 and Nc-1SAG4c2.1, that express the bradyzoite stage-specific protein NcSAG4 in the tachyzoite stage. These clones have similar growth rates in vitro as the wild-type (WT) strain Nc-1. Studies in a cerebral mouse model of infection revealed a slightly lower rate of detection of the transgenic strains in brains during the chronic phase of infection. However, a pregnant mouse model of infection revealed a reduction in the virulence of the Nc-1SAG4c1.1 strain despite the same tachyzoite expression of NcSAG4 and a similar anti-NcSAG4 response displayed by mice inoculated with Nc-1 SAG4c1.1 or Nc-1 SAG4c2.1 parasites. This behaviour may be related to the reduced ability of the Nc-1SAG4c1.1 parasites to invade host cells, which was observed in in vitro assays. The apparent reduction in persistence and the high growth rate of the transgenic strains, together with their constitutive expression of the protein NcSAG4, may be useful features for future immunoprophylaxis trials based on a safe live attenuated vaccine.

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Effects of Neospora caninum Infection at Mid-Gestation on Placenta in a Pregnant Mouse Model

Journal of Parasitology ◽

10.1645/ge-2347.1 ◽

2010 ◽

Vol 96 (5) ◽

pp. 1017-1020 ◽

Cited By ~ 8

Author(s):

I. C. López-Pérez ◽

E. Collantes-Fernández ◽

S. Rojo-Montejo ◽

V. Navarro-Lozano ◽

V. Risco-Castillo ◽

...

Keyword(s):

Mouse Model ◽

Neospora Caninum ◽

Pregnant Mouse ◽

Caninum Infection

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Broad human and animal coronavirus neutralisation by SARS-CoV-2 S2-targeted vaccination

10.1101/2021.11.30.470568 ◽

2021 ◽

Author(s):

Kevin W. Ng ◽

Nikhil Faulkner ◽

Katja Finsterbusch ◽

Mary Wu ◽

Ruth Harvey ◽

...

Keyword(s):

Immune System ◽

Severe Acute Respiratory Syndrome ◽

Mouse Model ◽

Antibody Response ◽

Common Cold ◽

Cross Reactivity ◽

Antibody Responses ◽

History Of

AbstractSeveral common-cold coronaviruses (HCoVs) are endemic in humans and several variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have emerged during the current Coronavirus disease 2019 (COVID-19) pandemic. Whilst antibody cross-reactivity with the Spike glycoproteins (S) of diverse coronaviruses has been documented, it remains unclear whether such antibody responses, typically targeting the conserved S2 subunit, contribute to or mediate protection, when induced naturally or through vaccination. Using a mouse model, we show that prior HCoV-OC43 S immunity primes neutralising antibody responses to otherwise subimmunogenic SARS-CoV-2 S exposure and promotes S2-targeting antibody responses. Moreover, mouse vaccination with SARS-CoV-2 S2 elicits antibodies that neutralise diverse animal and human alphacoronaviruses and betacoronaviruses in vitro, and protects against SARS-CoV-2 challenge in vivo. Lastly, in mice with a history of SARS-CoV-2 Wuhan-based S vaccination, further S2 vaccination induces stronger and broader neutralising antibody response than booster Wuhan S vaccination, suggesting it may prevent repertoire focusing caused by repeated homologous vaccination. The data presented here establish the protective value of an S2-targeting vaccine and support the notion that S2 vaccination may better prepare the immune system to respond to the changing nature of the S1 subunit in SARS-CoV-2 variants of concern (VOCs), as well as to unpredictable, yet inevitable future coronavirus zoonoses.

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In VitroandIn VivoEffects of the Bumped Kinase Inhibitor 1294 in the Related Cyst-Forming Apicomplexans Toxoplasma gondii and Neospora caninum

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01236-15 ◽

2015 ◽

Vol 59 (10) ◽

pp. 6361-6374 ◽

Cited By ~ 34

Author(s):

Pablo Winzer ◽

Joachim Müller ◽

Adriana Aguado-Martínez ◽

Mahbubur Rahman ◽

Vreni Balmer ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Vertical Transmission ◽

Kinase Inhibitor ◽

Neospora Caninum ◽

Antigen Expression ◽

Pregnant Mouse ◽

Content Type ◽

Elevated Expression ◽

Bumped Kinase Inhibitor

ABSTRACTWe report on thein vitroeffects of the bumped kinase inhibitor 1294 (BKI-1294) in cultures of virulentNeospora caninumisolates Nc-Liverpool (Nc-Liv) and Nc-Spain7 and in two strains ofToxoplasma gondii(RH and ME49), all grown in human foreskin fibroblasts. In these parasites, BKI-1294 acted with 50% inhibitory concentrations (IC50s) ranging from 20 nM (T. gondiiRH) to 360 nM (N. caninumNc-Liv), and exposure of intracellular stages to 1294 led to the nondisjunction of newly formed tachyzoites, resulting in the formation of multinucleated complexes similar to complexes previously observed in BKI-1294-treatedN. caninumbeta-galactosidase-expressing parasites. However, such complexes were not seen in a transgenicT. gondiistrain that expressed CDPK1 harboring a mutation (G to M) in the gatekeeper residue. InT. gondiiME49 andN. caninumNc-Liv, exposure of cultures to BKI-1294 resulted in the elevated expression of mRNA coding for the bradyzoite marker BAG1. Unlike in bradyzoites, SAG1 expression was not repressed. Immunofluorescence also showed that these multinucleated complexes expressed SAG1 and BAG1 and the monoclonal antibody CC2, which binds to a yet unidentified bradyzoite antigen, also exhibited increased labeling. In a pregnant mouse model, BKI-1294 efficiently inhibited vertical transmission in BALB/c mice experimentally infected with one of the two virulent isolates Nc-Liv or Nc-Spain7, demonstrating proof of concept that this compound protected offspring from vertical transmission and disease. The observed deregulated antigen expression effect may enhance the immune response during BKI-1294 therapy and will be the subject of future studies.

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