Differential Regulation of IL-9-Expression after Infection with Leishmania major in Susceptible and Resistant Mice

doi:10.1016/S0171-2985(11)80414-6

Immunobiology

Volume 189, Issue 5, December 1993, Pages 419-435

https://doi.org/10.1016/S0171-2985(11)80414-6 Get rights and content

Abstract

IL-9 is a pleiotropic lymphokine, one of its activities being the growth stimulation of certain CD4⁺ T lymphocytes. In murine cutaneous leishmaniasis, depending on the genetic background of the host mouse strain, vigorous proliferation of either mainly Th1 in resistant C57BL/6 mice or Th2-type CD4⁺ T cells in susceptible BALB/c mice occurs after infection with Leishmania major (L. major). Since little is known about the involvement of IL-9, the possible role of this cytokine with regard to its immunregulatory function was evaluated by comparing its presence in the serum and its expression kinetics in spleen and lymph nodes in resistant and susceptible mice.

To this sera of L. major-infected mice were tested functionally for IL-9. In addition the PCR-aided detection of IL-9 mRNA in organs of mice and measurement of the lymphokine in supernatants of restimulated lymph node and spleen cell cultures were used. We show here that although no functionally active IL-9 was detected in sera of both BALB/c and C57BL/6 mice, IL-9 is produced after in vitro antigenic restimulation and its mRNA was found to be expressed in lymph nodes and spleens during an immune response against L. major. Shortly after infection no principal differences in the kinetics of IL-9 expression could be observed, which had its maximum between day 5 and 7 after infection. The rate of production however was higher in the susceptible BALB/c mice. In athymic BALB/c nu/nu mice and in mice depleted of CD4⁺ T cells no IL-9 production was detectable in vivo at the level of mRNA and no IL-9 was produced after stimulation with L. major antigen in vitro. Treatment of infected mice with cyclosporin A ablates antigen-specific IL-9 production when tested in vitro without affecting its production after polyclonal T cell stimulation. Positively selected, purified CD4⁺ T cells were fully capable of producing IL-9. From 4 weeks after infection, IL-9 synthesis was observed oniy in BALB/c mice, correlating with the expansion of antigen-specific Th2 type T helper cells in these mice. Treatment of BALB/c mice with neutralizing anti-IL-4 mAb, a regimen known to lead to subsequent cure of infected BALB/c mice, suppressed late IL-9 synthesis.

References (37)

F.Y. Liew
Functional heterogeneity of CD4⁺ T cells in leishmaniasis
Immunol. Today.
(1989)
H.M. Wagner et al.
Interferon-γ inhibits the efficacy of interleukin 1 to generate a Th2-cell biased immune response induced by Leishmania major
Immunobiol.
(1991)
M.S. Krug et al.
First strand cDNA synthesis primed with oligo (dT)
Methods Enzymol.
(1987)
G. Trinchieri
Biology of natural killer cells
Adv. Immunol.
(1989)
J. Van Snick et al.
Cloning and characterization of cDNA for a new mouse T cell growth factor (P40)
J. Exp. Med.
(1989)
J.-C. Renauld et al.
Human P40/IL-9
Expression in activated CD4⁺ T cells, genomic organization, and comparison with the mouse gene. J. Immunol.
(1990)
L. Hültner et al.
Mast cell growth-enhancing activity (MEA) is structurally related and functionally identical to the novel mouse T cell growth factor P40/TCGF III (interleukin 9)
Eur. J. Immunol.
(1990)
J. Moeller et al.
Purification of MEA, a mast cell growth-enhancing activity to apparent homogeneity and its partial amino acid sequencing
J. Immunol.
(1990)
T. Suda et al.
Tumor necrosis factor alpha and P40 induce day 15 murine fetal thymocyte proliferation in combination with IL-2
J. Immunol.
(1990)
C. Uyttenhove et al.
Functional and structural characterization of P40, a mouse glycoprotein with T-cell growth factor activity
Proc. Natl. Acad. Sci. USA
(1988)

C. Uyttenhove et al.

Autonomous growth and tumorigenicity induced by P40/interleukin 9 cDNA transfection of a mouse P40-dependent T cell line

J. Exp. Med.

(1991)

E. Schmitt et al.

TCGF III/P40 is produced by naive murine CD4⁺ T cells but is not a general T cell growth factor

Eur. J. Immunol.

(1989)

C. Druez et al.

Functional and biochemical characterization of mouse P40/IL-9 receptors

J. Immunol.

(1990)

J.G. Howard

Immunological regulation and control of experimental leishmaniasis

Int. Rev. Exp. Pathol

(1986)

P. Scott et al.

Role of cytokines and CD4⁺ T-cell subsets in the regulation of parasite immunity and disease

Immunol. Rev.

(1989)

W.H. Boom et al.

Patterns of cytokine secretion in murine leishmaniasis: correlation with disease progression or resolution

Infect. Immun.

(1990)

R.L. Coffman et al.

The role of cytokines in the differentiation of CD4⁺ T cell subsets in vivo

Immunol. Rev.

(1991)

R.G. Tyws et al.

Therapeutic effect of anti-L3T4 monoclonal antibody GK1.5 on cutaneous leishmaniasis in genetically susceptible BALB/c mice

J. Immunol.

(1985)

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Polyamines produced by both prokaryotes and eukaryotes are bioactive substances with pleiotropic effects. Accumulating evidence has demonstrated that polyamines contribute to anti-inflammatory responses by suppressing the expression of proinflammatory cytokines in mononuclear cells and macrophages. However, the effects of polyamines on CD4⁺ T cell responses remain to be elucidated. Here, we investigated the effect of polyamines on cell fate decisions of naïve CD4⁺ T cells in vitro. We found that endogenously generated polyamines are essential for the development of T helper 2 (Th2) cells. Treatment with DL-2-difluoromethylornithine (DFMO), an inhibitor of polyamine biosynthesis, diminished GATA3 expression in CD4⁺ T cells under Th2-skewed conditions. Supplementation of exogenous polyamines rescued GATA3 downregulation caused by DFMO treatment in CD4⁺ T cells. Transcriptome analysis revealed that deprivation of endogenous polyamines resulted in upregulated Th9-related genes, such as Il9, Irf4, and Batf3, even under the Th2-skewing conditions. Depletion of intracellular polyamines reduced GATA3 expression but increased IL-9-producing CD4⁺ T cells under both Th2 and Th9-skewing conditions. Furthermore, oral administration of DFMO increased IL-9-producing CD4⁺ T cells in small intestine in mice. Thus, our data indicate that polyamines play a critical role in the regulation of the Th2/Th9 balance.
Interleukin-9 protects from early podocyte injury and progressive glomerulosclerosis in Adriamycin-induced nephropathy
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A wide spectrum of immunological functions has been attributed to Interleukin 9 (IL-9), including effects on the survival and proliferation of immune and parenchymal cells. In recent years, emerging evidence suggests that IL-9 expression can promote tissue repair in inflammatory conditions. However, data about the involvement of IL-9 in kidney tissue protection is very limited. Here, we investigated the role of IL-9 in Adriamycin-induced nephropathy (AN), a mouse model for proteinuric chronic kidney disease. Compared to wild type mice, IL-9 knockout (Il9^−/−) mice with AN displayed accelerated development of proteinuria, aggravated glomerulosclerosis and deterioration of kidney function. At an early stage of disease, the Il9^−/− mice already displayed a higher extent of glomerular podocyte injury and loss of podocyte number compared to wild type mice. In the kidney, T cells and innate lymphoid cells produced IL-9. However, selective deficiency of IL-9 in the innate immune system in Il9^−/−Rag2^−/− mice that lack T and B cells did not alter the outcome of AN, indicating that IL-9 derived from the adaptive immune system was the major driver of tissue protection in this model. Mechanistically, we could show that podocytes expressed the IL-9 receptor in vivo and that IL-9 signaling protects podocytes from Adriamycin-induced apoptosis in vitro. Finally, in vivo treatment with IL-9 effectively protected wild type mice from glomerulosclerosis and kidney failure in the AN model. The detection of increased serum IL-9 levels in patients with primary focal and segmental glomerulosclerosis further suggests that IL-9 production is induced by glomerular injury in humans. Thus, IL-9 confers protection against experimental glomerulosclerosis, identifying the IL-9 pathway as a potential therapeutic target in proteinuric chronic kidney disease.
Th9 cells are subjected to PD-1/PD-L1-mediated inhibition and are capable of promoting CD8 T cell expansion through IL-9R in colorectal cancer
2020, International Immunopharmacology
Th9 cells are named after their expression of IL-9. Studies in recent years demonstrated that Th9 cells could contribute to antitumor immunity by enhancing the recruitment and activation of mast cells, natural killer cells, CD8 T cells, and dendritic cells in the tumor microenvironment. To determine whether Th9 cells participate in colorectal cancer (CRC), we collected resected tumor samples from 20 CRC patients. In the tumor-infiltrating lymphocytes (TILs), IL-9⁺IL-4⁻ CD4⁺ T cells could be observed and were present at higher frequencies than the IL-9⁺IL-4⁺ and the IL-9⁻IL-4⁺ cells, suggesting that the majority of IL-9-producing TILs were bona fide Th9 cells. IL-9-secreting TILs presented particularly high PD-1 expression directly ex vivo. The expression of IL-9 was significantly reduced with PD-L1-mediated inhibition, which in turn was suppressed by anti-PD-1 blocking. Interestingly, the circulating CD4⁺ T cell compartment in CRC patients also presented Th9 enrichment, characterized by higher IL-9⁺IL-4⁻ and IL-9⁺IL-4⁺ cell frequencies in the CXCR3⁻CCR6⁻ compartment as compared to that in non-cancer controls. Using exogenous TGF-β and IL-4, we were capable of enriching Th9 cells without concurrent enrichment of Th2 cells. Th9-enriched CD4⁺ T cells, but not Th9-non-enriched cells, significantly increased the expansion of activated CD8⁺ T cells, in a manner that was dependent on the expression of IL-9R. In addition, the frequencies of Th9 cells in the tumor were positively correlated with the frequencies of CD8⁺ TILs. Together, we demonstrated that Th9 cells infiltrated CRC tumor, could be regulated via the PD-1/PD-L1 pathway, and could contribute the CD8⁺ T cell expansion.
Zinc supplementation plays a crucial role in T helper 9 differentiation in allogeneic immune reactions and non-activated T cells
2018, Journal of Trace Elements in Medicine and Biology
T helper (Th) 9 cells play a critical role in immune-mediated diseases, including allergic airway inflammation, autoimmune diseases, and cancer development. Thus, the promotion or suppression of Th9 cell differentiation, transcriptional control, and function is very important for a healthy immune system. Interestingly, T cell maturation, differentiation and function are highly dependent on the individuals’ zinc status. This is especially seen in zinc deficient individuals as in the elderly population often suffering of autoimmunity and increased incidence of infections.
In this regard, this study examines the impact of zinc supplementation in pharmacological doses on Th9 differentiation in two in vitro models: 1) in mixed lymphocyte cultures (MLC) displaying allogeneic activated T cells in graft versus host disease, and 2) on non-activated resting T cells in peripheral blood mononuclear cells (PBMC).
On the one hand, zinc supplementation significantly diminishes IL‐4-induced Th9 differentiation in MLC thereby ameliorating this pro-inflammatory allogeneic immunoreaction. On the other hand, Th9 cells are induced in resting T cells in PBMC hence triggering the immunological defense.
Thus, zinc supplementation can be considered as useful additive to dampen unwanted allogeneic immunoreactions. Moreover, the pro-inflammatory immune defense in non-reactive T cells can be strengthened, which is a frequent issue in the elderly population having a weakened immune response against invading pathogens.
IL-9 exhibits elevated expression in osteonecrosis of femoral head patients and promotes cartilage degradation through activation of JAK-STAT signaling in vitro
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Osteonecrosis of the femoral head (ONFH) often causes severe symptoms in young people and limits the mobility of the hip joint. Interleukin-9 (IL-9) is a multi-functional inflammatory factor that participates in lumbar disk herniation and arthritis and has been reported in many studies. However, the correlation between IL-9 and ONFH is unclear. The present study aimed to determine the role of IL-9 in the pathogenetic mechanism of osteonecrosis. To assess IL-9 expression in ONFH and femoral neck fracture patients, cartilage tissue was examined through western blot analysis and immunohistochemistry. Human primary chondrocytes were stimulated with IL-9, and inflammation-related cytokines and cartilage matrix-degrading enzymes were assessed via real-time PCR. After being treated with IL-9, Janus kinase-signal transducers and activators of transcription (JAK-STAT) signaling were tested through western blot analysis. Our results showed a significant increase in the expression of IL-9 in ONFH patients. IL-9 raised the level of inflammation-related cytokines and cartilage matrix-degrading enzymes and enhanced the activation of JAK-STAT signaling. Furthermore, blocking the JAK-STAT signaling pathway reduced the secretion of inflammation-related cytokines and cartilage matrix-degrading enzymes and markedly alleviated the degradation of the cartilage matrix. These findings provide new insights into the role that IL-9 plays in the pathogenetic mechanism of osteonecrosis and also provide a potential treatment for ONFH.
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Citation Excerpt :
It has also been reported that MAPK and insulin receptor substrate – PI3K pathways are activated through IL-9R signaling and play key roles in cell growth, survival, and differentiation.21,22 IL-9 was first thought to be a TH2-specific cytokine,23,24 but was later shown to be predominantly produced by a distinct subset of T helper cells, termed “TH9”.3 TH2, TH17, Tregs, and TH9 are now considered as some of the main IL-9-producing cells within the immune system.18,23,25,26
T helper 9 (T_H9) cells are considered as newly classified helper T cells that have an important role in the regulation of immune responses. Since these cells preferentially produce IL-9, these cells are termed T_H9 cells. Recently, the role of T_H9 and its signature cytokine (IL-9) has been investigated in a wide range of diseases, including autoimmunity, allergy, infections, cancer and immunodeficiency. Herein, we review the most recent data concerning T_H9 cells and IL-9 as well as their roles in disease. These insights suggest that T_H9 cells are a future target for therapeutic intervention.

View all citing articles on Scopus

: This work was supported by the Wilhelm Sander-Stiftung (grant 90.049.1).

^a: Dr. Dr. André Gessner, Institut für Klinische Mikrobiologie der Universität Erlangen-Nürnberg, Wasserturmstraße 3, 91054 Erlangen, Germany

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Differential Regulation of IL-9-Expression after Infection with Leishmania major in Susceptible and Resistant Mice

Abstract

Immunol. Today.

Immunobiol.

Methods Enzymol.

Adv. Immunol.

Cloning and characterization of cDNA for a new mouse T cell growth factor (P40)

J. Exp. Med.

Human P40/IL-9

Expression in activated CD4+ T cells, genomic organization, and comparison with the mouse gene. J. Immunol.

Mast cell growth-enhancing activity (MEA) is structurally related and functionally identical to the novel mouse T cell growth factor P40/TCGF III (interleukin 9)

Eur. J. Immunol.

Purification of MEA, a mast cell growth-enhancing activity to apparent homogeneity and its partial amino acid sequencing

J. Immunol.

Tumor necrosis factor alpha and P40 induce day 15 murine fetal thymocyte proliferation in combination with IL-2

J. Immunol.

Functional and structural characterization of P40, a mouse glycoprotein with T-cell growth factor activity

Proc. Natl. Acad. Sci. USA

Autonomous growth and tumorigenicity induced by P40/interleukin 9 cDNA transfection of a mouse P40-dependent T cell line

J. Exp. Med.

TCGF III/P40 is produced by naive murine CD4+ T cells but is not a general T cell growth factor

Eur. J. Immunol.

Functional and biochemical characterization of mouse P40/IL-9 receptors

J. Immunol.

Immunological regulation and control of experimental leishmaniasis

Int. Rev. Exp. Pathol

Role of cytokines and CD4+ T-cell subsets in the regulation of parasite immunity and disease

Immunol. Rev.

Patterns of cytokine secretion in murine leishmaniasis: correlation with disease progression or resolution

Infect. Immun.

The role of cytokines in the differentiation of CD4+ T cell subsets in vivo

Immunol. Rev.

Therapeutic effect of anti-L3T4 monoclonal antibody GK1.5 on cutaneous leishmaniasis in genetically susceptible BALB/c mice

J. Immunol.

Expression in activated CD4⁺ T cells, genomic organization, and comparison with the mouse gene. J. Immunol.

TCGF III/P40 is produced by naive murine CD4⁺ T cells but is not a general T cell growth factor

Role of cytokines and CD4⁺ T-cell subsets in the regulation of parasite immunity and disease

The role of cytokines in the differentiation of CD4⁺ T cell subsets in vivo