|IFN-γ||C57BL/6 (WT) and various KO mouse strains||N. caninum tachyzoites (Nc-1)||CD8−/− mice infected with N. caninum showed higher parasitic loads in the brain and lungs than WT ones.|
Mice treated with IFN-γ-expressing CD8+ T cells showed lower parasitic burdens than IFN-γ-deficient CD8+ T cells.
|IFN-γ and TNF-α||C57BL/6 mice||N. caninum tachyzoites (Nc-1)||Expression levels of IFN-γ and TNF-α were high in the brains of infected mice.|
The level of neurotransmitters glutamate, glycine, gamma-aminobutyric acid, dopamine and 5-hydroxytryptamine were altered in infected mice.
|IFN-γ and leptin||C57BL/6 mice||N. caninum tachyzoites (Nc-1)||In early infection, parasites were detected in the adipose tissue associated with increased numbers of immune cells, and increased expression of IFN-γ in gonadal adipose tissue. In chronic cases, parasite DNA was not detected in these tissues, but Th1 cell numbers remained above control levels, and marked increase of serum leptin was detected.|||
|IFN-γ||C57BL/6 mice ||N. caninum tachyzoites (Nc-1)||NK cells and various T cell populations mediate production of IFN-γ in the adipose tissue of N. caninum infected mice.|||
|IFN-γ and IL-12||BALB/c mice||N. caninum tachyzoites (Nc-1)||Mice treated with anti-IFN-γ alone increased morbidity/mortality, and increased foci of liver necrosis and increased parasite numbers in the lung by 7 dpi.|
Mice treated with rIL-12 decreased encephalitis and brain parasite load at 3 wpi.
|IFN-γ||BALB/c and IFN-γ−/− mice||N. caninum tachyzoites (Nc-1)||Infected IFN-γ−/− mice died earlier than WT. IFN-γ−/− mice failed to increase MHC class II expression on macrophages. BALB/c mice induced T-cell proliferation while IFN-γ−/− mice did not.|
In serum, high levels of IFN-γ and IL-4 were detected in resistant hosts, whereas IL-10 was detected in IFN-γ−/− mice. The levels of IL-12 in IFN-γ−/− mice were higher than in BALB/c mice at 7 dpi.
|IFN-γ||BALB/3T3 clone A31 mice||N. caninum tachyzoites (Nc-1)||The viability of N. caninum-infected murine fibroblast cells was significantly reduced after treatment with mouse IFN-γ. FasL expression was clearly induced by N. caninum-infection and IFN-γ- treatment, and the reduction in host-cell viability was prevented with the addition of anti-mouse FasL monoclonal antibody (mAb).|||
|IFN-γ and IL-4||BALB/c and IFN-γ−/− mice||Tachyzoites and SRS2 antigen N. caninum (Nc-1)||In the acute infection of N. caninum, IFN-γ−/− mice showed high levels of IL-10 production, whereas significant levels of IFN-γ and IL-4 production were observed in resistant WT mice. |
BALB/c mice vaccinated with a virus expressing NcSRS2 were protected against parasite and low levels of IFN-γ and high levels of IL-4 productions were observed.
|IFN-γ and IL-12||A/J mice||N. caninum tachyzoite and antigen (Nc-1)||Inbred A/J mice developed no clinical and little histologic evidence of infection by N. caninum. Splenocytes obtained from infected mice proliferate in vitro in response to both N. caninum-soluble antigens. Mice infected with N. caninum produce significant quantities of IL-12 and IFN-γ.|||
|IFN-γ||CBA/Ca and Swiss white (BK: W) mice||N. caninum tachyzoites (Nc-1)||Infected spleen cells had the highest specific lymphoproliferative response, and a mixed cytokine response with elevated IFN-γ and fairly low IL-4 and IL-10 secretion was recorded.|||
|IFN-γ, TNF-α, IL-10 and TGF-β,||Rat ||N. caninum tachyzoites (Nc-1)||Treatment of primary mixed cultures of rat astrocytes and microglia with either IFN-γ or TNF-α reduced infection rate.|
In the absence of IL-10 and TGF-β, tachyzoite numbers were reduced significantly against non-treated cells.
|IFN-γ, TNF-α, IL-17A and IL-4||Fat-tailed dunnart||N. caninum tachyzoites (Nc-Nowra) ||mRNA expression during the time course of infection revealed higher levels of IFN-γ, TNF-α, IL-17A, and IL-4 cytokines in infected rather than non-infected dunnart spleen cells.|||
|IFN-γ and TNF-α||Cattle||N. caninum tachyzoites (Nc-1)||Addition of recombinant IFN-γ in primary astroglia-microglia culture inhibited the tachyzoite growth, which was not reversed by the addition of an NO antagonist. TNF-α, to a lesser extent, also inhibited the tachyzoite growth.|||
|IFN-γ, IL-12, TNF-α, IL-10||Heifers ||N. caninum tachyzoites(Illinois cattle isolate)||Infected dams showed an increased number of lymphocyte subpopulations compared with uninfected pregnant animals.|
Gene expression increased both Th1 and Th2 cytokines in N. caninum-infected animals (IFN-γ, IL-12, TNF-α, IL-10).
|IFN-γ||Cattle||N. caninum tachyzoites (Nc-Liv)||The live and heat-inactivated tachyzoites of N. caninum, directly trigger production of IFN-γ from purified and IL-2-activated bovine NK cells.|||
|IFN-γ and IL-4 ||Cattle||N. caninum tachyzoites (Nc-Liv)||Percentages of CD2+ and CD4+ T-cells in peripheral blood mononuclear cells (PBMC) increased after infection in both early and late gestation.|
Percentages of CD8+ T-cells increased 1–2 wpi at day 70. IL-4 and IFN-γ mRNA expression in PBMC increased 1–2 wpi at day 210 and IL-4 increased 1–2 wpi at day 70.
|IL-2, IFN-γ, IL-12p40, TNF-α, IL-18, IL-10, and IL-4||Heifer||N. caninum tachyzoites (Nc-Liv)||Infection in early gestation induced an increase in mRNA levels of IL-2, IFN-γ, IL-12p40, TNF-α, IL-18, IL-10, and IL-4 in placental tissues.|
This was associated with extensive placental necrosis and an infiltration of CD4+ T cells and macrophages, and IFN-γ and TGF-β was also highly and moderately increased, respectively.
|IFN-γ and IL-17||Cattle||N. caninum tachyzoites (Nc-Liv)||Naive T-cells in contact with infected macrophages produced both IFN-γ and IL-17 in a pattern that is dependent on whether the priming macrophage was protected or non-protected.|||
|IFN-γ, IL-4, IL-12p40, IL-10, TNF-α and MHC class II||Heifer||N. caninum-naturally infected animals||An increase in IFN-γ and IL-4 mRNA was detected. IL-12p40, IL-10, and TNF-α were also significantly increased.|
MHC Class II antigens were expressed on maternal and fetal epithelial and stromal fibroblastoid cells.
|IFN-γ, TNF-α, IL-12p40, IL-10, and IL-4||Heifer||N. caninum-naturally infected animals||N. caninum infected dams showed elevated mRNA levels of IFN-γ, TNF-α and IL-12p40, and IL-10, but expression of IL-4 did not differ significantly among the groups.|| |
|IFN-γ, IL12, TNF-α, IL-6, IL-10 and IL-4||Heifer||N. caninum tachyzoites (Illinois cattle isolate)||Fetuses had higher expression of most cytokines at 3 and 9 wpi in fetuses that were alive at 6 wpi.|
In dams, most cytokines were down-regulated from 6 wpi, with elevated IL-4 expression observed in the caruncle.
|IFN-γ, IL-10 and IL-4||Heifer||N. caninum tachyzoites (Nc-Spain7)||In infected dams with live fetuses, IFN- γ increased in both caruncle and cotyledon, and IL-10 elevated in cotyledon.|
Infected live fetuses showed elevated expression of IFN-γ and IL-10 in fetal spleen, and showed diminished expression of IL-4 in the thymus compared to control uninfected fetuses.
|IFN-γ, IL-17 and IL-4||Heifer||N. caninum infection (Nc-Spain7)||In dams, significantly higher IFN-γ and IL-4 levels were found in the experimentally infected animals compared to the control or naturally infected dams.|
IL-17A production was very low in the dams infected with N. caninum and did not seem to be a major regulator of IFN-γ production in this model.
|IFN-γ||Sheep||N. caninum tachyzoites (Nc-1 and Nc-Liv)||The N. caninum Nc-1 multiplied more quickly in fibroblast cells than the Nc-Liv isolate.|
Treatment of the cells with ovine rIFN-γ for 24h before infection inhibited intracellular multiplication of the parasite.
|IFN-α, IFN-β and IFN-γ||Dog||N. caninum tachyzoites (Nc-1)||IFN-γ inhibited the parasite growth in MDCK cells infected with N. caninum tachyzoites with greater efficacy than IFN-α and IFN-β.|||
|IFN-γ||Dog and BALB/c cell lines||N. caninum tachyzoites (Nc-1)||In the presence of IFN-γ, the viability of the infected host cell was decreased and apoptotic cell death occurred. An increase of FasL expression on the IFN-γ-treated cells following N. caninum infection was observed.|
IFN-γ treatment decreased Bcl-2 expression in the cells cultured with N. caninum while parasite infection increased Bcl-2.
|TNF-α and IL-12||BALB/c mice ||N. caninum cytoplasmic|
dynein LC8 light chain (NcDYNLL) protein (Nc-1)
|NcDYNLL2 is a secretory protein and was internalized by the host immune cells and stimulated TNF-α and IL-12 production by murine dendritic cells.|||
|N. caninum tachyzoites (Nc-1)||The number of splenic conventional dendritic cells (cDCs) increased at 5 dpi, while the number of plasmacytoid dendritic cells (pDCs) did not change on infection, this effect is associated with upregulation of costimulatory and MHC class II molecules.|
This stimulatory effect was more marked at the earliest assessed time point after infection, 12 h, when a clear increase in the frequency of cDCs and pDCs producing IL-12 was also observed.
|IFN-γ,TNFR2, IL-10, beta 2 microglobulin (b2M), and inducible nitric oxide synthase (iNOS2)||Various KO and WT mice||N. caninum tachyzoites (virulent Nc-1 or Nc-2 or attenuated Ncts-8)||Infection with Nc-1 was 100% lethal in IFN-γ−/− mice. TNFR2−/− and b2M−/− mice were infected with Nc-1 or Nc-2 isolate, while TNFR2−/− or b2M−/− mice were resistant to Ncts-8 infection.|
Lack of mortality and minimal histopathology scores demonstrated that Nc-1, Nc-2, and Ncts-8 infections were avirulent in IL-10 and iNOS2−/− mice.
|TNF-α and NO||Rat||N. caninum tachyzoites (Nc-Bahia isolate)||Astrocytes responded to infection by producing the pro-inflammatory cytokine TNF-α and the neurotoxic-free radical NO, 24, and 72 hpi.|||
|iNOS||C57BL/6 mice and iNOS−/− mice||Neospora tachyzoite (Nc-1)||N. caninum infection in WT mice induced NO production in vivo and in vitro, and iNOS−/− mice succumbed during acute infection, associated with increased in parasite load.|
Infected iNOS−/− mice showed marked CNS inflammation, and increased production of IL-12, IFN-γ, IL-6, TNF-α, and IL-17A.
|GRO-a, IL-8 and IP-10, MCP-1, RANTES, GM-CSF, COX-2 and iNOS||Cattle||N. caninum tachyzoites|
|N. caninum infection in the bovine umbilical vein endothelial cells (BUVEC) in vitro resulted in elevation of the CXC chemokines GRO-a, IL-8, and IP-10, the CC chemokines MCP-1 and RANTES and of GM-CSF, COX-2, and iNOS after 2–4 hpi.|||
|IL-17||Cattle||N. caninum tachyzoites (Nc-Liv)||Infection with the N. caninum induced fibroblasts to secrete pro-IL-17 factors by inducing a γΔ17 phenotype that preferentially kills infected target cells.|||
|TNF-α and IL-8||Cattle||N. caninum tachyzoites (Nc-Spain7 and Nc-Spain1H)||TNF-α and IL-8 were elevated, while IL-6 and TGF-β1 were decreased in both trophoblast and caruncular cell lines.|
Higher secretion of TNF-α were noticed in the trophoblast cell line infected with the low-virulence Nc-Spain1H.
|Delayed type hypersensitivity (DTH)||Cattle ||N. caninum infection followed by intradermal inoculation of soluble tachyzoite antigen (Nc-1)||Either experimental or natural infection of cows with live N. caninum tachyzoites developed skin reactions compatible with DTH between 24 and 96 hpi associated with an increase in IFN-γ release.|||
|NO and advanced oxidation protein products (AOPP)||Goat||N. caninum-naturally infected animals||N. caninum seropositive animals showed higher serum levels of NO compared to seronegative animals.|
AOPP levels did not change in infected and non-infected groups.
|IL-4||Qs mice||N. caninum tachyzoites (Nc-Liv and Nc-SweB1)||Spleen cells from both infected/non-pregnant mice produced higher IFN-γ, IL-12, and TNF-α than in infected/pregnant mice.|
IL-4 was exclusively increased in infected/pregnant mice and thus appear to be responsible for the observed decline in Th1 cytokine production in pregnant mice.
|IL-4||BALB/c mice||N. caninum tachyzoites (Nc-1)||In naïve mice before pregnancy, neutralization of IL-4 during gestational challenge did not result in decreased congenital transmission, while in mice that were primed before pregnancy, congenital transmission was significantly decreased.|
Decreased congenital transmission was associated with significantly lower levels of maternal splenocyte IL-4 secretion, lower IL-4 mRNA levels, and higher levels of IFN-γ secretion.
|Neutrophil extracellular trap||Cattle ||N. caninum tachyzoites (Nc-1)||N. caninum tachyzoites triggered NETosis in a time- and dose-dependent manner. NET structures are released by bovine PMN and entrapping tachyzoites.|||
|Monocyte extracellular trap (ETs)||Goat||N. caninum tachyzoites (Nc-1)||N. caninum tachyzoites were not only capable of triggering ETs formation in caprine monocytes, but also that monocyte-released ETs were able to entrap viable tachyzoites.|||
|Neutrophils extracellular traps (NETs)||Dog||N. caninum tachyzoites (Nc-1)||N. caninum tachyzoite induced NETs formation as observed by scanning electron microscopy.|||
|Indoleamine 2,3-Dioxygenase (IDO)||Cattle||N. caninum tachyzoites (Nc-1)||Induction of the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO) inhibited the parasite growth that is mediated by IFN-γ-activated bovine fibroblasts and endothelial cells.|||
|MIF, TGF-β, IFN-γ and IDO||Human cell lines||N. caninum tachyzoites (Nc-1)||N. caninum infection increased the macrophage migration inhibitory factor (MIF), mainly in HeLa cells. Conversely, parasite infection induced down-regulation of TGF-β, mostly in BeWo cells.|
HeLa cells were more susceptible to Neospora infection than BeWo cells mediated by IFN-γ via IDO-dependent pathways in HeLa cells alone.
|iNOS, IDO and COX-2||Rats||N. caninum tachyzoites (Nc-Bahia)||iNOS, IDO, and COX-2 control the proliferation of N. caninum in vitro, while the release of IL-10 by glia affects the inflammation and maintains the parasitism.|||
|Perforin and Granzyme||Cattle ||N. caninum tachyzoites (Nc-1)||Enrichment and blocking of CD4+ and CD8+ T-lymphocyte effector subsets indicated that CD4+ CTL killed N. caninum-infected, autologous target cells and that killing was mediated through a perforin/granzyme pathway.|||
|Lectin ||Heifers||N. caninum tachyzoites (Nc-1)||Changes in the lectin-binding pattern were noted in infected animals as noticed in the glycocalyx, apical cytoplasm of endometrial cells, and apical cytoplasm of the trophoblastic cells.|||
|Pregnancy-associated glycoprotein; PAG-1, PAG-2||Heifers||N. caninum tachyzoites (Nc-Spain7) ||Non-aborting infected heifers showed a temporary fall in PAG-1 and PAG-2 concentrations from 7 to 14 dpi.|
Dams aborting at 14 and 21 dpi showed dramatic PAG-1 and PAG-2 decreases from 14 dpi to undetectable levels upon euthanasia.
|SERPINA14||Heifers||N. caninum tachyzoites (Nc-Spain7)||Normally, uterine serpins (SERPINA14) regulate the immunosuppressive effect of progesterone during late pregnancy.|
N. caninum infection downregulates the uterine immunosuppressive function of SERPINA14.
|SERPINA14||Heifers||N. caninum tachyzoites (Nc-Spain7)||Intercaruncular SERPINA14 expression was negatively correlated with IFN-γ expression in cotyledon samples and with IL4 expression in uterine lymph nodes.|||
|Neurotrophic factors GFAP, BDNF and NGF and cytokine IL-10,||Rat||N. caninum tachyzoites (Nc-1)||N. caninum-infected glial cultures responded with astrogliosis increased GFAP, IL-10, BDNF, and NGF gene expression.|||
|Host nutrient molecules||Human and bovine cell lines||N. caninum tachyzoites (Nc-Liv) ||N. caninum utilizes plasma lipoproteins, scavenges cholesterol from Niemann–Pick type C protein 1 -containing endocytic organelles, and salvages sphingolipids from host Golgi Rab14 vesicles.|||
|Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE)||Gerbils||N. caninum tachyzoites (Nc-1)|
|On 7 dpi a decrease of AChE in total blood and brain was observed, along with reduction of BChE in plasma of infected animals compared to noninfected. |
AChE activity increased in total blood and reduced in brain at 30 dpi, and BChE activity was markedly increased at 30 dpi.
|Purine||Gerbils ||N. caninum tachyzoites (Nc-1)||The purine levels (ATP, ADP, AMP, adenosine, inosine and xanthine) in the brain are markedly reduced at 7 dpi, while the purine levels were significantly increased on days 15 (ATP, AMP, adenosine, hypoxanthine, and xanthine) and on 30 PI (ATP, ADP, AMP, adenosine, and uric acid).|||
|Cathelicidin||Human cell line||N. caninum tachyzoites (Nc-1)||Infection of macrophages (THPI) with live tachyzoites elevated cathelicidins were associated with increased pro-inflammatory cytokines (TNFα, IL-1β, IL-8, IFN-γ). This immune response in infected macrophages was mediated by (MEK 1/2) and resulted in reduced parasite internalization in naïve macrophages.|||