Immune response evaluation in Balb/c mice after crude extract of Anisakis typica sensitization

Background and Aim: Anisakis is a global challenge for a fish product which may lead to a decrease in economic value and consumers' preference. Skipjack (Katsuwonus pelamis) in Kupang, Nusa Tenggara Timur, Indonesia, have important economic value for local fisheries. Anisakis typica is one of the Anisakis species which potent to induce an allergic reaction. However, the study about A. typica involved in the dendritic cells (DCs), T helper 1 (Th1), T helper 2 (Th2), and regulatory T cells (Tregs) is still limited. This study aimed to analyze the dynamic changed of the immune system including DCs, CD4+ T cells, and Tregs after 1 week of A. typica sensitization. Materials and Methods: Twenty-four male Balb/C mice were randomly divided into four groups (n=6), mice treated with crude A. typica extract (CAE) 50, 75, and 100 mg/kg BW, respectively. CAE was given orally per day for a week. At the end of the experiment, the animals were sacrificed and the spleen was collected. DCs were labeled as CD11c+ interleukin-6+ (IL-6+); CD4+ T cells were distinguished as Th1 (CD4+ interferon-γ+ [IFN-γ+]) and Th2 (CD4+ IL-4+ and CD4+ IL-5+); Tregs were labeled as CD4+CD25+CD62L+. The expression of each cell was determined by flow cytometry. Results: Our result described that CAE elicits CD11c+ IL-6+, CD4+ IFN-γ+, CD4+ IL-4+, and CD4+ IL-5+ and reduces CD4+CD25+CD62L+ significantly (p<0.05) in dose-dependent manner in mice after A. typica infection. Conclusion: The Th1/Th2 ratio after A. typica crude extract treatment exhibits a mixed pattern rather than the classical model allergy to food antigens. Our study is expected as a basic understanding of the changes in immune response after A. typica infection.

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Hydrophilic Astragalin Galactoside Induces T Helper Type 1-Mediated Immune Responses via Dendritic Cells

International Journal of Molecular Sciences ◽

10.3390/ijms19103120 ◽

2018 ◽

Vol 19 (10) ◽

pp. 3120

Author(s):

Jae Jeon ◽

Byung-Cheol Lee ◽

Doman Kim ◽

Daeho Cho ◽

Tae Kim

Keyword(s):

T Cells ◽

Immune Responses ◽

Cd4 T Cells ◽

Biological Activities ◽

Water Soluble ◽

Th1 Cells ◽

Dependent Manner ◽

T Helper ◽

Th1 Cell ◽

Ifn Γ

A flavonoid Astragalin (kaempferol-3-O-β-d-glucopyranoside, Ast) has several biological activities including anti-oxidant, anti-HIV, and anti-allergic effects. Nonetheless, its insolubility in hydrophilic solvents imposes restrictions on its therapeutic applications. In this study, we investigated the effects of water-soluble astragalin-galactoside (kaempferol-3-O-β-d-isomaltotrioside, Ast-Gal) on murine bone marrow-derived dendritic cell (DC) maturation and T helper (Th) cell-mediated immune responses. Ast-Gal significantly increased maturation and activation of DCs through the upregulation of surface markers, such as cluster of differentiation (CD)80, CD86, and Major histocompatibility complex (MHC) II in a dose-dependent manner, while Ast had little effects. Additionally, Ast-Gal-treated DCs markedly secreted immune-stimulating cytokines such as interleukin (IL)-1β, IL-6, and IL-12. Importantly, Ast-Gal strongly increased expression of IL-12, a polarizing cytokine of Th1 cells. In a co-culture system of DCs and CD4+ T cells, Ast-Gal-treated DCs preferentially differentiates naïve CD4+ T cells into Th1 cells. The addition of neutralizing IL-12 monoclonal antibody (mAb) to cultures of Ast-Gal-treated DCs and CD4+ T cells significantly decreased interferon (IFN)-γ production, thereby indicating that Ast-Gal-stimulated DCs enhance the Th1 response through IL-12 production by DCs. Injection with Ast-Gal-treated DCs in mice increased IFN-γ-secreting Th1 cell population. Collectively, these findings indicate that hydrophilically modified astragalin can enhance Th1-mediated immune responses via DCs and point to a possible application of water-soluble astragalin-galactoside as an immune adjuvant.

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Hydrophilic Astragalin Galactoside Induces T Helper Type 1-Mediated Immune Responses via Dendritic Cells

10.20944/preprints201808.0357.v1 ◽

2018 ◽

Author(s):

Jae Hyoung Jeon ◽

Byung-Cheol Lee ◽

Doman Kim ◽

Daeho Cho ◽

Tae Sung Kim

Keyword(s):

T Cells ◽

Immune Responses ◽

Cd4 T Cells ◽

Biological Activities ◽

Water Soluble ◽

Th1 Cells ◽

Dependent Manner ◽

T Helper ◽

Th1 Cell ◽

Ifn Γ

A flavonoid Astragalin (kaempferol-3-O-β-D-glucopyranoside, Ast) has several biological activities including anti-oxidant, anti-HIV, and anti-allergic effects. Nonetheless, its insolubility in hydrophilic solvents imposes restrictions on its therapeutic applications. In this study, we investigated the effects of water-soluble astragalin-galactoside (kaempferol-3-O- β-D-isomaltotrioside, Ast-Gal) on dendritic cell (DC) maturation and T helper (Th) cell-mediated immune responses. Ast-Gal significantly increased maturation and activation of DCs through up-regulation of surface markers, such as CD80, CD86, and MHC II in a dose-dependent manner, while Ast had little effects. Also, Ast-Gal-treated DCs markedly secreted immune-stimulating cytokines such as IL-1β, IL-6, and IL-12. Importantly, Ast-Gal strongly increased expression of IL-12, a polarizing cytokine of Th1 cells. In a co-culture system of DCs and CD4+ T cells, Ast-Gal-treated DCs preferentially differentiates naïve CD4+ T cells into Th1 cells. The addition of neutralizing IL-12 mAb to cultures of Ast-Gal-treated DCs and CD4+ T cells significantly increased IFN- γ production, thereby indicating that Ast-Gal-stimulated DCs enhance the Th1 response through IL-12 production by DCs. Injection with Ast-Gal-treated DCs in mice increased IFN-γ-secreting Th1 cell population. Collectively, these findings indicate that hydrophilically modified astragalin can enhance Th1-mediated immune responses via DCs, and point to a possible application of water-soluble astragalin-galactoside as an immune adjuvant.

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The effect of Kefir on The Immune Response of Healthy Volunteers In Vitro

Journal of Agromedicine and Medical Sciences ◽

10.19184/ams.v3i2.5067 ◽

2017 ◽

Vol 3 (2) ◽

pp. 28

Author(s):

Desie Dwi Wisudanti

Keyword(s):

Immune Response ◽

T Cells ◽

Immune Responses ◽

Healthy Volunteers ◽

Cd8 T Cells ◽

Cd4 T Cells ◽

Fermented Milk ◽

Bioactive Components ◽

Ifn Γ

Kefir is a functional foodstuff of probiotics, made from fermented milk with kefir grains containing various types of beneficial bacteria and yeast. There have been many studies on the effects of oral kefir on the immune system, but few studies have shown the effect of bioactive components from kefir (peptides and exopolysaccharides/ kefiran), on immune responses. The purpose of this study was to prove the effect of kefir supernatant from milk goat on healthy immune volunteer response in vitro. The study was conducted on 15 healthy volunteers, then isolated PBMC from whole blood, then divided into 5 groups (K-, P1, P2, P3 and P4) before culture was done for 4 days. The harvested cells from culture were examined for the percentage of CD4+ T cells, CD8+ T cells, IFN-γ, IL-4 using flowsitometry and IL-2 levels, IL-10 using the ELISA method. The results obtained that kefir do not affect the percentage of CD4+ T cells and CD8+ T cells. The higher the concentration of kefir given, the higher levels of secreted IFN- γ and IL-4, but a decrease in IL-2 levels. Significant enhancement occurred at levels of IL-10 culture PBMC given kefir with various concentrations (p <0.01), especially at concentrations of 1%. These results also show the important effects of kefir bioactive components on immune responses. The conclusion of this study is that kefir can improve the immune response, through stimulation of IL-10 secretion in vitro.

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Polarization of Allogeneic T-Cell Responses by Influenza Virus-Infected Dendritic Cells

Journal of Virology ◽

10.1128/jvi.74.17.7738-7744.2000 ◽

2000 ◽

Vol 74 (17) ◽

pp. 7738-7744 ◽

Cited By ~ 25

Author(s):

Sangkon Oh ◽

Maryna C. Eichelberger

Keyword(s):

Immune Response ◽

T Cells ◽

Dendritic Cells ◽

Influenza Virus ◽

T Cell ◽

Interleukin 2 ◽

Dependent Manner ◽

Ifn Γ ◽

High Doses ◽

Type Response

ABSTRACT The developing immune response in the lymph nodes of mice infected with influenza virus has both Th1- and Th2-type characteristics. Modulation of the interactions between antigen-presenting cells and T cells is one mechanism that may alter the quality of the immune response. We have previously shown that the ability of dendritic cells (DC) to stimulate the proliferation of alloreactive T cells is changed by influenza virus due to viral neuraminidase (NA) activity. Here we show that DC infected with influenza virus A/PR/8/34 (PR8) stimulate T cells to produce different types of cytokines in a dose-dependent manner. Optimal amounts of the Th1-type cytokines interleukin-2 (IL-2) and gamma interferon (IFN-γ) were produced from T cells stimulated by DC infected with low doses of PR8, while the Th2-type cytokines IL-4 and IL-10 were produced only in response to DC infected with high doses of PR8. IL-2 and IFN-γ levels corresponded with T-cell proliferation and were dependent on the activity of viral NA on the DC surface. In contrast, IL-4 secretion required the treatment of T cells with NA. Since viral particles were released only from DC that are infected with high doses of PR8, our results suggest that viral NA on newly formed virus particles desialylates T-cell surface molecules to facilitate a Th2-type response. These results suggest that the activity of NA may contribute to the mixed Th-type response observed during influenza virus infection.

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Tofacitinib modulates the VZV-specific CD4+ T cell immune response in vitro in lymphocytes of patients with rheumatoid arthritis

Rheumatology ◽

10.1093/rheumatology/kez175 ◽

2019 ◽

Vol 58 (11) ◽

pp. 2051-2060 ◽

Cited By ~ 10

Author(s):

Giovanni Almanzar ◽

Felix Kienle ◽

Marc Schmalzing ◽

Anna Maas ◽

Hans-Peter Tony ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Granzyme B ◽

Janus Kinase ◽

Cell Immune Response ◽

Dependent Manner ◽

Th1 Response

AbstractObjectiveRA is a chronic inflammatory disease characterized by lymphocyte infiltration and release of inflammatory cytokines. Previous studies have shown that treatment with Janus kinase inhibitors, such as tofacitinib, increased the incidence rate of herpes zoster compared with conventional DMARDs. Therefore, this study aimed to investigate the effect of tofacitinib on the varicella-zoster-virus (VZV)-specific T cell immune response.MethodsThe effect of tofacitinib on the VZV-specific T cell immune response was determined by evaluating the IFNγ production, the proliferative capacity, the VZV-induced differentiation into effector and memory T cells, the expression of activation marker CD69 and helper T cell type 1 (Th1)-characteristic chemokine receptors, such as CXCR3 and CCR5, as well as cytotoxic activity (perforin and granzyme B expression) of CD4+ T cells of patients with RA compared with healthy donors upon stimulation with VZV antigen in vitro.ResultsTofacitinib significantly reduced the IFNγ production, proliferation, activation, and CXCR3 expression of VZV-specific CD4+ T cells in a dose-dependent manner in short- and long-term lymphocyte culture. No effect on the distribution of naive, effectors or memory, or on the expression of perforin or granzyme B by VZV-specific CD4+ T cells was observed.ConclusionThis study showed that tofacitinib significantly modulated the Th1 response to VZV. The poor VZV-specific cellular immune response in patients with RA may be considered in recommendations regarding appropriate vaccination strategies for enhancing the VZV-specific Th1 response.

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Influenza virus-specific CD4+ T helper type 2 T lymphocytes do not promote recovery from experimental virus infection.

Journal of Experimental Medicine ◽

10.1084/jem.180.4.1273 ◽

1994 ◽

Vol 180 (4) ◽

pp. 1273-1282 ◽

Cited By ~ 196

Author(s):

M B Graham ◽

V L Braciale ◽

T J Braciale

Keyword(s):

Immune Response ◽

T Cells ◽

T Lymphocytes ◽

Cd4 T Cells ◽

Primary Role ◽

T Helper ◽

Helper Type

T lymphocytes play a primary role in recovery from viral infections and in antiviral immunity. Although viral-specific CD8+ and CD4+ T cells have been shown to be able to lyse virally infected targets in vitro and promote recovery from lethal infection in vivo, the role of CD4+ T lymphocytes and their mechanism(s) of action in viral immunity are not well understood. The ability to further dissect the role that CD4+ T cells play in the immune response to a number of pathogens has been greatly enhanced by evidence for more extensive heterogeneity among the CD4+ T lymphocytes. To further examine the role of CD4+ T cells in the immune response to influenza infection, we have generated influenza virus-specific CD4+ T cell clones from influenza-primed BALB/c mice with differential cytokine secretion profiles that are defined as T helper type 1 (Th1) clones by the production of interleukin 2 (IL-2) and interferon gamma (IFN-gamma), or as Th2 clones by the production of IL-4, IL-5, and IL-10. Our studies have revealed that Th1 clones are cytolytic in vitro and protective against lethal challenge with virus in vivo, whereas Th2 clones are noncytolytic and not protective. Upon further evaluation of these clonal populations we have shown that not only are the Th2 clones nonprotective, but that pulmonary pathology is exacerbated as compared with control mice as evidenced by delayed viral clearance and massive pulmonary eosinophilia. These data suggest that virus-specific CD4+ T cells of the Th2 subset may not play a primary role in virus clearance and recovery and may lead to immune mediated potentiation of injury.

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Cyclophosphamide modulates CD4+ T cells into a T helper type 2 phenotype and reverses increased IFN-γ production of CD8+ T cells in secondary progressive multiple sclerosis

Journal of Neuroimmunology ◽

10.1016/j.jneuroim.2003.10.036 ◽

2004 ◽

Vol 146 (1-2) ◽

pp. 189-198 ◽

Cited By ~ 35

Author(s):

Arnon Karni ◽

Konstantin Balashov ◽

Wayne W. Hancock ◽

Padmanabhan Bharanidharan ◽

Michal Abraham ◽

...

Keyword(s):

Multiple Sclerosis ◽

T Cells ◽

Cd8 T Cells ◽

Cd4 T Cells ◽

Progressive Multiple Sclerosis ◽

T Helper ◽

Secondary Progressive ◽

Ifn Γ ◽

Helper Type

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Adaptive Immune Responses Mediated Age-related Plasmodium yoelii 17XL and 17XNL Infections in 4 and 8-week-old BALB/c Mice

10.21203/rs.2.15639/v1 ◽

2019 ◽

Author(s):

Qiu-bo Wang ◽

Yun-ting Du ◽

Fei Liu ◽

Xiao-dan Sun ◽

Xun Sun ◽

...

Keyword(s):

T Cells ◽

Survival Rate ◽

Cd4 T Cells ◽

Plasmodium Yoelii ◽

Th2 Cells ◽

Dependent Manner ◽

Adaptive Immune ◽

Age Related ◽

Ifn Γ ◽

Old Mice

Abstract Background As the quest to eradicate malaria continues, it is important to clarify the opposite clinical outcomes between childhood and adulthood. The relationship between adaptive immune response and age-related malaria infection remains unknown. Methods 4 and 8-week-old mice were used to mimic childhood and adulthood, respectively. Parasitemia and the survival rate were monitored. The proportion and function of Th1 and Th2 cells were detected by FACS. The levels of IFN-γ, IL-4, total IgG, IgG1, IgG2a and Plasmodium yoelii MSP-1-special IgG were measured by ELISA. Results Infant mice were more susceptible to P. yoelii 17XNL infection, with lower survival rate and higher parasitemia. The adult group showed greater resistance to P. yoelii 17XL infection, with lower parasitemia. Compared with 4-week-old mice, the percentage of CD4 + T-bet + IFN-γ + Th1 cells as well as IFN-γ production were significantly increased on day 5 p.i. in the 8-week-old mice after P. yoelii 17XNL infection. The percentage of CD4 + GATA3 + IL-4 + Th2 cells and CD4 + CXCR5 + Tfh cells, and IL-4 production in the 8-week-old mice obviously increased on day 5 and day 10 after P. yoelii 17XNL infection. Notably, the levels of total IgG, IgG1, IgG2a and P. yoelii MSP-1-special IgG were also significantly increased in the 8-week-old mice. PD-1, a marker of exhaustion, was up-regulated on CD4 + or activated CD4 + T cells in the 8-week-old mice as compared to the 4-week-old group. Conclusion We consider that enhanced cellular and humoral adaptive immunity might contribute to rapid clearance of malaria among adults, likely in a PD-1-dependent manner due to induction of CD4 + T cells exhaustion.

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Anti-Leukemia Immune Responses in CML Patients Treated with Imatinib Mesylate.

Blood ◽

10.1182/blood.v106.11.1108.1108 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1108-1108

Author(s):

Christiane I.-U. Chen ◽

Holden T. Maecker ◽

Wesley H. Neal ◽

Rhoda Falkow ◽

Peter P. Lee

Keyword(s):

Immune Response ◽

T Cells ◽

Immune Responses ◽

Cd4 T Cells ◽

Post Treatment ◽

Leukemia Cells ◽

Tnf Α ◽

Ifn Γ ◽

Cytogenetic Remission ◽

Pre Treatment

Abstract Imatinib mesylate, a selective inhibitor of the bcr/abl tyrosine kinase, has revolutionized the treatment of patients with chronic myelogenous leukemia (CML). Most CML patients in chronic phase achieve hematologic remission with imatinib, while some achieve cytogenetic remission. As imatinib is an oral agent with few side effects, it has rapidly become the first-line therapy for most CML patients. However, this therapy does not represent a cure, as patients who discontinue the drug invariably relapse. Furthermore, imatinib resistance is beginning to emerge in some patients. Hence, the need to find alternate, potentially curative, therapies for CML remains. To date, the only curative treatment for CML is allogeneic bone marrow or stem cell transplantation (ABMT). A major mechanism of the curative potential of ABMT is immunological, as evidenced by the poor clinical outcome with T cell-depleted ABMT, and the efficacy of donor lymphocyte infusions (DLI) upon relapse. We hypothesized that an effective anti-leukemia immune response may emerge in patients entering remission on imatinib which may contribute to its clinical effectiveness. If so, strategies to further enhance this anti-leukemia immune response may lead to a potential cure. To determine if CML patients in remission on imatinib develop anti-leukemia immune responses, blood and bone marrow samples from patients before and after treatment were collected and analyzed. Pre-treatment samples were utilized as sources of autologous leukemic cells to detect anti-leukemia immune responses in post-treatment samples in IFN-g ELISPOT assays. Pre-treatment samples alone, post-treatment samples alone, and when available, serial post-treatment samples mixed together served as controls. In 9 of 14 patients investigated, IFN-g release was detected in pre- and post-treatment samples together with a median response of 22 spots above background (range 10 – 56 dots, p<0.01), whereas serial post-treatment samples together in 8 patients yielded results similar to background (median 5, range 5 – 20). In 6 of these patients in hematologic (or cytogenetic) remission, sufficient cells were available to allow additional analyses via intracellular staining for IFN-g, TNF-a, and IL-2 in autologous leukemia stimulated T cells (CD4 and CD8) and NK cells. In 4 of 6 patients, leukemia-reactive T cells were detected, most prominently in CD4+ T cells expressing TNF-a (1.4 – 37%), followed by IL-2 (0.3 – 12%) and IFN-g (0.1 – 4.6%). NK cells did not show significant expression of these cytokines upon stimulation with autologous leukemia cells. In pre-treatment and post-treatment samples alone, IL-2, TNF-a, and IFN-g expression was not detectable (0 – 0.5%). These results suggest that a significant portion of CML patients in remission with imatinib develop an anti-leukemia immune response, most notably in CD4+ T cells. Mechanisms by which imatinib treatment leads to anti-leukemia immune responses, and the molecular targets to which these cells are directed, will be further investigated. This knowledge will be useful in the development of immunotherapy strategies against CML as well as other leukemias, and raises the hope that immunotherapy may be combined with imatinib to eradicate residual leukemia cells for a durable cure of the disease. intracellular cytokine staining CD4+ T Cells CD8+ T Cells IL-2 IFN- γ TNF- α IL-2 IFN- γ TNF- α pt 1 0.3 0 0.8 0.1 0.1 0.5 pt 1 0.3 0.1 1.4 0.1 0.1 0.4 pt 2 2.6 0.8 10.3 2.2 2.1 6.1 pt 3 21 2 37 2.3 0.7 1.7 pt 4 12 4.6 19 6.3 1.8 5.8

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NFAT1 in Adult and Cord Blood-Derived Human CD4+ T-Cells: Regulatory Implications for Graft vs. Host Disease (GVHD).

Blood ◽

10.1182/blood.v108.11.3164.3164 ◽

2006 ◽

Vol 108 (11) ◽

pp. 3164-3164

Author(s):

R.P. Weitzel ◽

M.L. Lesnewski ◽

Y. Huang ◽

L.R. Fanning ◽

Mary Laughlin

Keyword(s):

T Cells ◽

Cord Blood ◽

Lower Incidence ◽

Cd4 T Cells ◽

Dependent Manner ◽

Cd4 Cells ◽

Rt Pcr ◽

Host Disease ◽

Ifn Γ

Abstract Clinical benefits of umbilical cord blood (UCB) grafts include the lower incidence of acute graft-versus-host disease (aGVHD) following allogeneic transplantation. We have previously demonstrated dramatically lower expression of the Nuclear Factor of Activated T-Cells 1 (NFAT1) protein, its associated transcripts and known binding partners, but not NFAT1 transcripts in UCB CD4+ cells following primary stimulation when compared to T-cells obtained from adult blood (AB). As NFAT1 is a key transcriptional regulator of immune responses, we hypothesize that NFAT1 and NFAT1-dependent factors play a critical role in the increased proliferation and decreased cytokine production seen in UCB, which may contribute to the lower incidence of aGVHD following UCB transplantation observed clinically. We have now demonstrated in vitro via siRNA-mediated knockdown and subsequent analysis of adult-derived primary T-cells with RT-PCR and Cytometric Bead Assay (CBA) that expression of a GVHD-promoting, Th1-type cytokine profile is dependent on expression of NFAT1, and that knockdown of NFAT1 results in a transcriptional and cytokine secretion profile similar to that seen in UCB T-cells with respect to key immunoregulatory cytokines and signal transducers following stimulation. Specifically, siRNA-mediated reduction of NFAT1 by at least 68% results in 93% and 28% reduction of IFN-γ, and TNF-αrespectively, as well as a 55% reduction in CTLA4 measured by RT-PCR. Additionally, short hairpin RNA (shRNA)-mediated stable knockdown of NFAT1 results in an enhanced and prolonged reduction of IFN-γ, IL-2, and TNF-αsecretion over transient siRNA transfection in both the Jurkat cell line and adult primary CD4+ T-cells. However, microarray and RT-PCR time point analyses indicate NFAT1 transcripts are not changed in UCB versus AB CD4+ cells. Under siRNA knockdown conditions, we observed at least a 24-hour delayed decrease in NFAT1 protein levels following mRNA reduction and an eventual rebound of both NFAT1 and its associated cytokines following transient transfection at 48 hours. This suggests a crucial regulatory role for post-translational events which may preferentially occur in UCB-derived T-cells in vivo. Indeed, we have observed ubiquitination of NFAT1 and rapid rescue of NFAT1 expression following treatment with the calpain-proteasome inhibitor Acetyl-L-Leucyl-L-Leucyl-L-Norleucinal (ALLN) in UCB-derived T-cells in vitro by co-immunoprecipitation (co-IP) in a dose-dependent manner. NFAT1 in unstimulated UCB CD4+ cells is shown by co-IP to be significantly more ubiquitinated than in AB-derived cells. Overall, our data indicates that regulation of NFAT1 through ubiquitination and proteasomal degradation, and its subsequent downstream effects may constitute a key difference between T-cells of adult and neonatal origin; specifically that this post-translational modification may serve as a method by which UCB-derived T-cells may prevent NFAT1 activity, impacting their allogeneic response.

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