Interleukin-35 on B cell and T cell induction and regulation
Huang et al. Journal of Inflammation
Interleukin-35 on B cell and T cell induction and regulation
Ai Huang 0 1 3
Lin Cheng 0 2
Miao He 1 3
Jun Nie 1 3
Jianjun Wang 1 3
Ke Jiang 1 3
0 Equal contributors
1 Department of Thoracic Surgery, Union hospital, Tongji Medical College, Huazhong University of Science and Technology , No.1277 Jiefang Avenue, Wuhan, Hubei Province 430022 , People's Republic of China
2 Department of Anesthesiology, Union hospital, Tongji Medical College, Huazhong University of Science and Technology , No.1277 Jiefang Avenue, Wuhan, Hubei Province 430022 , People's Republic of China
3 Department of Thoracic Surgery, Union hospital, Tongji Medical College, Huazhong University of Science and Technology , No.1277 Jiefang Avenue, Wuhan, Hubei Province 430022 , People's Republic of China
Interleukin (IL)-35 is a relatively newly discovered member of IL-12 cytokine family that is unique in that it is a dimer formed by two subunits. The review documents the structure, secretion and signal transduction of IL-35, the regulation effect of IL-35 on B cells and T cells as well as the adoptive transfer of IL-35+ regulatory B cells (Breg), therapeutic prospects of recombinant IL-35 (rIL-35) and IL-35 regulation role in various diseases. B-cell regulation expands the regulatory range of IL-35 and alters the view that IL-10 is the chief immune mechanism for Breg cells which secrete IL-35. IL-35 induces Breg cells, which then can induce Treg cells. IL-35 also plays an immunomodulatory role in the human body.
Interleukin −35; B cells; T cells
Interleukin-35 (IL-35) structure and secretion
IL-35 was found by Niedbala [
] and Collison [
simultaneously and it is reported to contain IL-12α chain
p35 and IL-27β chain Epstein-Barr virus-induced gene 3
(Ebi3) connected by disulfide bond. IL-35, initially named
at the 13th International Congress of Immunology, is the
new focus of cytokines research. IL-35 is similar to other
IL-12 family members which are heterodimeric
glycoproteins formed with disulfide-linked α (p19, p28, or p35)
and β (p40 or EBi3) chains. The α-chain has 4-α-helical
bundles, a typical cytokine structure, and the β-chain is
homologous to the soluble cytokine receptor. p35 and p40
combine to form IL-12; p19 and p40 combine to form
IL23; p28 and Ebi3 combine to form IL-27 [
]. IL-35 is
composed of p35 and Ebi3, and it differs from the expression
and secretion way of other IL-12 members. In response to
bacteria, bacterial products, or intracellular parasites,
IL12, IL-23, and IL-27 are secreted by activated
antigenpresenting cells, including B cells, monocyte, macrophages
and dendritic cells [
]. IL-35 was initially reported to be
produced by Treg cells and was essential for maximizing
the inhibitory role of Treg cells [
]. Recently studies
suggest that regulatory B cells (Breg) also produce IL-35 and
rIL-35 fusion proteins can induce Breg cells to secret
IL10 and IL-35 [
IL-35 receptors and signal transduction
Peptide chain sharing is common to the IL-12 family as
they bind to receptors to activate signal transducer and
activator of transcription (STAT) proteins [
]. IL-35 is
uniquely anti-inflammatory cytokine in contrast to other
IL-12 pro-inflammatory cytokines [
]. This difference is
thought to be associated with the receptors and signaling
pathways specific to IL-35 and future studies should
confirm these assertions. An obstacle to understanding
the molecular mechanism underlying IL-35 is the lack of
clarity about the IL-35 receptor (IL-35R) and its signal
transduction pathway [
]. Collison’s group reported that
mouse IL-35R differed from traditional cytokine receptors.
IL-35R is composed of dimers which are IL-12Rβ2
homodimers, gp130 homodimers or IL- IL-12Rβ2/gp130
]. IL-35 binds to IL-35R and initiates signal
transduction and exert biological function. IL-12Rβ2 or
gpl30 homodimer activates STAT4 or STAT1, but only
IL35R in the IL-12Rβ2/gp130 heterodimer form can induce
STAT1- and STAT4-activated signaling pathways to
mediate Treg cell function and positively or negatively feedback
regulate IL-35 gene expression [
]. IL-35 is reported to
activate STAT1 and STAT4 in T cells [
] where IL-35 binds
to gp130 and IL-12Rβ2 receptors [
], both of which
depend on JAK-STAT signaling to introduce intracellular
responses. However in B cells, IL-35 signaling mediates
STAT1 and STAT3 activation through IL-12Rβ2: IL-27Ra
] (Fig. 1).
IL-35 regulation and signal transduction in Breg cells
B cells have been traditionally thought to contribute to
immune defense by secreting antibodies and antigen
presentation but they also have function in immune
regulation as Breg cells [
]. There are multiple B cell
subsets which have immune regulation function, such as
CD138+ plasma cells [
], B10 cells (CD1dhiCD5hi) [
CD21hiCD23hiCD24hi transitional 2-marginal zone
precursor cells [
], and Tim-1+ B cells [
], but Breg cells
do not have a unified determined phenotype. It is
generally recognized that Breg cells play a role in
immunosuppressive function by secreting IL-10 under the
stimulation of toll-like receptor (TLR) agonists, CD40L,
and IL-21 [
Furthermore immunosuppressive pathways of Breg
cells may not solely depend on IL-10 [
] as research
suggests that Breg cells can produce IL-35 and that
rIL35 can induce Breg cells to secrete IL-10 and IL-35 [
]. B-cell-derived IL-35 also acts on T cell-induced
Foxp3+ Treg cells [
] (Fig. 2). Although rIL-35
inhibits B220hi B cell proliferation, it selectively induces
CD19+CD5+B220lo Breg cell proliferation in vivo [
Unlike T cells, inhibition of gp130 (with small interfering
RNA [siRNA] or neutralizing antibodies) does not affect
IL-35-mediated of B cell proliferation or IL-10 secretion.
In contrast, silence of IL-12Rß2 and IL-27Ra in B cells
completely blocks inhibition function of IL-35. Thus IL-35
may mediate biological activity in different cell types via
different receptors and STAT signaling pathways and
future studies are required to confirm those [
IL-35 regulation and signal transduction in T cells
IL-35 is a novel inhibitory cytokine that may be
specifically produced by Treg cells. It is required for maximal
suppressive activity of Treg. In addition, IL-35 can block
the proliferation of Th1 and Th17 cells by limiting early
T cell rest on the G1 phase of cell division [
Although IL-35 can inhibit Th1 proliferation, it is
resistant to Treg conversion due to the potent inhibition of
Ebi3 and p35 transcription by IFNγ from Th1. Moreover,
IL-35 blocks Th2 development by repressing GATA3
and IL-4 expression and limiting Th2 proliferation.
IL-35 can also mediate conversion of Th2 cells to Treg,
although this can be blocked by IFN-γ [
Like TGF-β and IL-10, IL-35 can induce the
development of an induced regulatory T cell (iTreg) population,
iTr35, which suppressed T-cell proliferation via
]. iTreg do not express Foxp3, IL-10, and
TGFβ (Fig. 3). iTr35 were as effective as nTregs at
restoring immune homeostasis and preventing
autoimmunity disease in Foxp3−/− mice by limiting the
proliferation of T cells and so prevented the
modeling of EAE and IBD and promoted the proliferation
of B16 tumor cells. Adoptive transfer IL-35
treatment increased the proliferation of Foxp3+CD39+
CD4+ T cells which secreted IL-10 for autoimmune
protection in a collagen-induced arthritis model [
However, whether the presence of IL-35 can mediate
Treg amplification under physiological conditions is
While gp130 is fairly ubiquitously expressed, IL-12Rβ2 is
expressed mainly on the surfaces of activated T cells,
natural killer cells, B cells, and dendritic cells [
]. IL-12Rβ2 is
undetectable on most resting T cells, but can be rapidly
upregulated by exposure to IL-2, IFN-γ, IL-12, IL-27, and
TNF-α. Indeed, IL-2 or IL-27 pretreatment increases T cell
sensitivity to IL-35 mediated suppression [
IL-35 mediates the inhibitory effect on T cells through
the signal pathway of STAT1 and STAT4, but also lead
to the pro-inflammatory effects by activating the STAT
molecule of IFN-γ and IL-12, in which the key difference
is that IL-35 induced STAT1-STAT4 heterodimer
Highly purified heterodimeric cytokine IL-35 is difficult to
obtain and this is a limitation to immunology research,
especially for elucidating the role of this cytokine in
autoimmune, infectious diseases and tumor immunity.
Transgenic technology has been applied to infect cells to
establish a rIL-35 mouse model in which rIL-35 is a
heterodimer of p35 and Ebi3 [
]. Similar to IL-27, IL-35 is not
secreted as a disulfide-linked heterodimer as Ebi3
associates non-covalently with the IL-12p35 [
]. The native
Ebi3/p35 heterodimer would be difficultly to isolate in vivo
because only about 4% of the secreted Ebi3 co-precipitated
with the IL-12p35 in vitro over expression studies in COS7
]. Meanwhile, as the absence of IL-12p35
substantial amount of the Ebi3 degraded in the ER (endoplasmic
reticulum), thereby reducing the bioavailability of ebi3.
These contribute to the low levels of IL-35 in vivo [
Others generated recombinant mouse IL-35 (rIL-35) using
a bicistronic vector containing IRES (internal ribosomal
entry site) that allowed stoichio etric expression of the Ebi3
and IL-12p35 [
]. Another approach that has been used is
to construct a heterodimeric protein covalently linking
Ebi3 and IL-12p35 [
IL-35regulation in various diseases
(1) IL-35 and autoimmune diseases.
Early studies mainly used a model of p35 or ebi3 deficient
mice. However, the absence of IL-12α affects the function of
IL-12 as well as that of IL-27 and IL-35. Despite these
complexities, Ebi3−/− and Il-12a−/− mice model also provide the
proof that IL-35 have immune suppressive function in some
autoimmune diseases such as experimental autoimmune
uveitis (EAU) [
], experimental allergic encephalomyelitis
], collagen-induced arthritis (CIA) [
], and other conditions. That IL-35 fusion protein
obtained from genetic engineering produced data consistent
with the knockout model results and studies indicates that
injection of rIL-35 can be used to protect against allergic
airway model [
] as well as colitis [
] and autoimmune
]. Studies show that IL-35 regulates immune
function via T cells in many diseases. In NOD transgenic
mice, infusion of IL-35 over expressing islet cells significantly
alleviated symptoms of diabetes and local inflammatory
responses, reduced the number of effector CD4+ and CD8+
T cells in pancreatic islet tissue, inhibited infiltration of T cells
from transitioning from G1 to S phases, and decreased
effector T cells proliferation [
]. In a type II CIA model,
infusion of rIL-35 reduced local inflammatory responses by
inhibiting Th17 cell differentiation. In an Ebi3 subunit
knockout mouse model, enhanced Th17 cell infiltration and
aggravated inflammatory response was noted [
To study a potential regulatory role of IL-35+ Breg, a
model of EAU was established and rIL-35 or IL-35+
Breg could control the incidence and development of
EAU. IL-35+ Breg cell-mediated protection depended on
induction and proliferation of endogenous Breg cells and
Foxp3+ Treg cells and inhibition of pathogenic Th1 and
Th17 effector cells [
]. Adoptive infusion of
rIL-35mediated Breg cells improved the animal health after
EAU eradication as well. This suggested that in vitro
study of functional rIL-35+ Breg cells may give insight
into potential roles for Breg and IL-35 Breg cells in
autoimmune disease and cancer. IL-35-derived Treg
cells (i.e., iTR35) can maintain Breg cells, but IL-35+
Breg cells can also induce Treg cells. Therefore, in an
inflammation-derived IL-35-dependent regulatory
environment, different cell populations generated by IL-35
may interact via positive feed forward mechanisms and
induce iTR35 and IL-35 + Breg cells through infectious
] (Fig. 4).
IL-35 and tumor immunity
Previous work suggests that IL-35 plays a role in tumor
immune escape. Ebi3 expression was increased in
Hodgkin’s lymphoma [
] and more Treg cells and
inhibitory cytokines were noted in peripheral blood and
tumor microenvironments of patients with pancreatic or
breast carcinomas [
]. Ebi3 expression in lung cancer
cells has also been found to be associated with tumor
progression and siRNA-mediated down-regulation of the
Eib3 gene inhibited proliferation of lung cancer cells
]. Similarly, inoculation of b16 tumor cells in
Ebi3knockout mice showed enhanced anti-tumor immunity
relative to wild-type mice and the metastatic potential of
tumor cells were suppressed. In an IL-35-positive tumor
cell microenvironment, studies showed that there were
significant increase in CD11b + Gr1+ myeloid-derived
suppressor cells (MDSC) and vascular endothelial
growth factors promoted tumor angiogenesis [
MDSCs were immunosuppressive and inhibited
cytotoxic T cells. In this way, tumor cell-generated IL-35
may protect against cytotoxic T-cell-mediated damage.
In contrast, applications of neutralizing antibody to IL-35
significantly inhibited tumor cell proliferation [
Although many studies have shown that IL-35
contributes to immunosuppressive function in the mouse model,
but this effect is limited in humans [
]. No expression of
IL-35-derived Treg cells had been observed in early
investigations in humans. This suggests that IL-35 may not be
constitutively expressed in humans. However, it can be
produced in specific tissues and cell types after a certain
inflammatory stimulus. Recent studies have shown that,
under strong stimulation, Treg can produce IL-35, and
CD4+ T cells under the stimulation of IL-35 can express
IL35similar to iTr35 in mice [
]. Similarly, in humans, CD8+
Treg can suppress the immune response against prostate
cancer by expressing CTLA-4 and IL-35 [
more research is required to draw any conclusions,
preliminary information suggests that elevated serum IL-35 is
associated with tumor malignancy [
] and clinical
stage and decreased IL-35 is associated with autoimmune
disease and chronic infection [
]. (Fig. 4).
IL-35 and infectious diseases
IL-35, as an inhibitory cytokine, plays an important role
in infectious diseases. The study suggested that
Mycobacterium tuberculosis (M. tuberculosis) could induce T
cell proliferation and foster IFN-γ production in
p35deficient mice, P40 deficient mice, and wild-type mice,
thus eliminating the pathogen. Mice lacking p40 have
been found to be less able to show antigen-specific
responses than those lacking the p35 subunit. However,
their ability to counteract infection is less pronounced
than that of wild-type mice [
]. In addition, the
protective responses can be induced in wild-type and
p35deficient mice by inoculating with vaccines, and
increased the secretion of IFN-γ and IL-17. However,
P40-deficient mice did not produce antigen-specific
IFN-γ or IL-17, and increased the infection load of
bacteria. In addition, treatment of p35-deficient mice
with Candida albicans (C. albicans) reduced the rate of
fungal infection and was associated with little to no
obvious symptoms of infection relative to P19-deficient
mice. In p35-deficient mice and those in which p35 gene
was disrupted, the immunosuppressive function of IL-35
was inhibited, which impaired anti-fungal immunity
]. The CD4+ T cells in the peripheral blood of
patients with chronic hepatitis B also showed high levels
of expression of p35 and EB13 protein, indicating that
IL-35 was related to the immune response of chronic
hepatitis B patients [
]. These results showed that,
during the acute infection process, IL-35 preferentially
activated Th1 cells, stimulated proliferation of Treg cells,
and inhibited the differentiation of Th17 cells, thereby
preventing excessive tissue damage caused by the
clearing of pathogens. In the chronic infection and
inflammation, IL-35 selectively inhibited effector cells, including
Th17 cells, which slowed down the development of
autoimmune diseases [
]. (Fig. 4).
B-cell regulation expands the regulatory range of IL-35
and alters the view that IL-10 is the chief immune
mechanism for Breg cells which secrete IL-35. IL-35
signaling mediates STAT1 and STAT3 activation in B
cells by binding to IL-12Rβ2 and IL-27Ra receptors and
inducing Treg cells which provide mutual stimulation
under inflammation and amplification of regulatory cells.
IL-35-induced B cells also transform to secretary IL-35+
Breg cells and rIL-35 or IL-35+ Breg cells control the
incidence and development of EAU. IL-35 induces Breg
cells, which then can induce Treg cells. In the case of
inflammation, cells which can generate IL-35 provide
mutual stimulation, resulting in amplification of
regulatory cells. Recent researchers have found that in some
inflammatory stimulus conditions, IL-35 also play an
immunomodulatory role in the human body, so we can
look forward to further exploring immunotherapy
approaches through IL-35.
Future studies may address whether IL-35 shares its
receptors with other IL-12 members and rIL-35 may
help clarify these biological effects and identify other cell
types or subtypes involved in immune regulation by
producing IL-35. How IL-35 inhibits cell proliferation,
why different cells mediate different signaling pathways,
and potential prospects of clinical use of IL-35 (rIL-35
and IL-35 Breg cells) as a chemotherapeutic or to treat
autoimmune diseases or organ transplantation also await
Breg: Regulatory B cells; CIA: Collagen-induced arthritis; EAU: Experimental
autoimmune uveitis; Ebi3: Epstein-Barr virus-induced gene 3; ER: endoplasmic
reticulum; IL-35: Interleukin-35; IL-35R: IL-35 receptor; iTR35: IL-35-derived Treg cells;
MDSC: Myeloid-derived suppressor cells; rIL-35: recombinant IL-35; siRNA: small
interfering RNA; STAT: Signal transducer and activator of transcription; TLR: Toll-like
receptor; Treg: Regulatory T cells
Availability of data and materials
AH and LC drafted the initial manuscript, MH, JN and JJW modified the
manuscript as submitted. KJ corrected/approved the final manuscript as
submitted. All authors read and approved the final manuscript.
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1. Niedbala W , Wei XQ , Cai B , Hueber AJ , Leung BP , IB MI. IL-35 is a novel cytokine with therapeutic effects against collagen-induced arthritis through the expansion of regulatory T cells and suppression of Th17 cells . Eur J Immunol . 2007 ; 37 : 3021 - 9 .
2. Collison LW , Workman CJ , Kuo TT , Boyd K , Wang Y , Vignali KM , et al. The inhibitory cytokine IL- 35 contributes to regulatory T- cell function . Nature . 2007 ; 450 : 566 - 9 .
3. Tedder TF , Leonard WJ . Autoimmunity:regulatory B cells-IL-35 and IL21regulate the regulators . Nat Rev Rheumatol . 2014 ; 10 : 452 - 3 .
4. Ma X , Trinchieri G . Regulation of interleukin-12 production in antigenpresenting cells . Adv Immunol . 2001 ; 79 : 55 - 92 .
5. O 'Shea JJ , Paul WE . Regulation of T (H) 1 differentiation-controlling the controllers . Nat Immunol . 2002 ; 3 : 506 - 8 .
6. Hunter CA. New IL-12-family members: IL-23 and IL-27, cytokines with divergent functions . Nat Rev Immunol . 2005 ; 5 : 521 - 31 .
7. Wang RX , Yu CR , Dambuza IM , Mahdi RM , Dolinska MB , Sergeev YV , et al. Interleukin -35 induces regulatory B cells that suppress autoimmune disease . Nat Med . 2014 ; 20 : 633 - 41 .
8. Shen P , Roch T , Lampropoulou V , O'Connor RA , Stervbo U , Hilgenberg E , et al. IL-35 -producing B cells are critical regulators of immunity during autoimmune and infectious diseases . Nature . 2014 ; 507 : 366 - 70 .
9. Collison LW , Delgoffe GM , Guy CS , Vignali KM , Chaturvedi V , Fairweather D , et al. The composition and signaling of the IL-35 receptor are unconventional . Nat Immunol . 2012 ; 13 : 290 - 9 .
10. Yanaba K , Bouaziz JD , Haas KM , Poe JC , Fujimoto M , Tedder TF , et al. A regulatory B cell subset with a unique CD1dhiCD5+ phenotype controls T cell-dependent inflammatory responses . Immunity . 2008 ; 28 : 639 - 50 .
11. Neves P , Lampropoulou V , Calderon-Gomez E , Roch T , Stervbo U , Shen P , et al. Signaling via the MyD88 adaptor protein in B cells suppresses protective immunity during salmonella typhimurium infection . Immunity . 2010 ; 33 : 777 - 90 .
12. Evans JG , Chavez-Rueda KA , Eddaoudi A , Meyer-Bahlburg A , Rawlings DJ , Ehrenstein MR , et al. Novel suppressive function of transitional 2 B cells in experimental arthritis . J Immunol . 2007 ; 178 : 7868 - 78 .
13. Ding Q , Yeung M , Camirand G , Zeng Q , Akiba H , Yagita H , et al. Regulatory B cells are identified by expression of TIM-1 and can be induced through TIM-1 ligation to promote tolerance in mice . J Clin Invest . 2011 ; 121 : 3645 - 56 .
14. Yoshizaki A , Miyagaki T , DiLillo DJ , Matsushita T , Horikawa M , Kountikov EI , et al. Regulatory B cells control T-cell autoimmunity through IL-21- dependent cognate interactions . Nature . 2012 ; 491 : 264 - 8 .
15. Wilson MS , Taylor MD , O'Gorman MT , Balic A , Barr TA , Filbey K , et al. Helminth-induced CD19+CD23hi B cells modulate experimental allergic and autoimmune inflammation . Eur J Immunol . 2010 ; 40 : 1682 - 96 .
16. Wirtz S , Billmeier U , Mchedlidze T , Blumberg RS , Neurath MF . Interleukin-35 mediates mucosal immune responses that protect against T-cell-dependent colitis . Gastroenterology . 2011 ; 141 : 1875 - 86 .
17. Vignali DA , Kuchroo VK . IL-12 family cytokines: immunological playmakers . Nat Immunol . 2012 ; 13 : 722 - 8 .
18. Collison LW , Chaturvedi V , Henderson AL , Giacomin PR , Guy C , Bankoti J , et al. IL -35-mediated induction of a potent regulatory T cell population . Nat Immunol . 2010 ; 11 : 1093 - 01 .
19. Presky DH , Yang H , Minetti LJ , Chua AO , Nabavi N , Wu CY , et al. A functional interleukin 12 receptor complex is composed of two beta-type cytokine receptor subunits . Proc Natl Acad Sci U S A . 1996 ; 93 : 14002 - 7 .
20. Canda-Sánchez A , Salgado FJ , Pérez-Díaz A , Varela-González C , Arias P , Nogueira M. Differential distribution of both IL-12Rbeta chains in the plasma membrane of human T cells . J Membr Biol . 2009 ; 227 : 1 - 12 .
21. Maldonado RA , Soriano MA , Perdomo LC , Sigrist K , Irvine DJ , Decker T , et al. Control of T helper cell differentiation through cytokine receptor inclusion in the immunological synapse . J Exp Med . 2009 ; 206 : 877 - 92 .
22. Devergne O , Birkenbach M , Kieff E. Epstein-Barr virus -induced gene 3 and the p35 subunit of interleukin 12 form a novel heterodimeric hematopoietin . Proc Natl Acad Sci U S A . 1997 ; 94 : 12041 - 6 .
23. Devergne O , Hummel M , Koeppen H , Le Beau MM , Nathanson EC , Kieff E , et al. A novel interleukin-12 p40-related protein induced by latent EpsteinBarr virus infection in B lymphocytes . J Virol . 1996 ; 70 : 1143 - 53 .
24. Aparicio-Siegmund S , Moll JM , Lokau J , Grusdat M , Schroder J , Plohn S , et al. Recombinant p35 from bacteria can form interleukin (IL-)12, but not IL-35 . PLoS One. 2014 ; 9 : e107990 .
25. Kochetkova I , Golden S , Holderness K , Callis G , Pascual DW . IL-35 stimulation of CD39+ regulatory T cells confers protection against collagen II-induced arthritis via the production of IL-10 . J Immunol . 2010 ; 184 : 7144 - 53 .
26. Tirotta E , Duncker P , Oak J , Klaus S , Tsukamoto MR , Gov L , et al. Epstein-Barr virus-induced gene 3 negatively regulates neuroinflammation and T cell activation following coronavirus-induced encephalomyelitis . J Neuroimmunol . 2013 ; 254 : 110 - 6 .
27. Huang CH , Loo EX , Kuo IC , Soh GH , Goh DL , Lee BW , et al. Airway inflammation and IgE production induced by dust mite allergen-specific memory/effector Th2 cell line can be effectively attenuated by IL-35 . J Immunol . 2011 ; 187 : 462 - 7 .
28. Bettini M , Castellaw AH , Lennon GP , Burton AR , Vignali DA . Prevention of autoimmune diabetes by ectopic pancreatic beta-cell expression of interleukin-35 . Diabetes. 2012 ; 61 : 1519 - 26 .
29. Niedobitek G , Päzolt D , Teichmann M , Devergne O . Frequent expression of the Epstein-Barr virus (EBV)-induced gene, EBI3, an IL-12 p40-related cytokine, in Hodgkin and reed-Sternberg cells . J Pathol . 2002 ; 198 : 310 - 6 .
30. Liyanage UK , Moore TT , Joo HG , Tanaka Y , Herrmann V , Doherty G , et al. Prevalence of regulatory T cells is increased in peripheral blood and tumor microenvironment of patients with pancreas or breast adenocarcinoma . J Immunol . 2002 ; 169 : 2756 - 61 .
31. Nishino R , Takano A , Oshita H , Ishikawa N , Akiyama H , Ito H , et al. Identification of Epstein-Barr virus-induced gene 3 as a novel serum and tissue biomarker and a therapeutic target for lung cancer . Clin Cancer Res . 2011 ; 17 : 6272 - 86 .
32. Wang Z , Liu JQ , Liu Z , Shen R , Zhang G , Xu J , et al. Tumor-derived IL-35 promotes tumor growth by enhancing myeloid cell accumulation and angiogenesis . J Immunol . 2013 ; 190 : 2415 - 23 .
33. Bardel E , Larousserie F , Charlot-Rabiega P , Coulomb-L'Herminé A , Devergne O. Human CD4+ CD25+ Foxp3+ regulatory T cells do not constitutively express IL-35 . J Immunol . 2008 ; 181 : 6898 - 05 .
34. Seyerl M , Kirchberger S , Majdic O , Seipelt J , Jindra C , Schrauf C , et al. Human rhinoviruses induce IL-35-producing Treg via induction of B7-H1 (CD274) and sialoadhesin (CD169) on DC . Eur J Immunol . 2010 ; 40 : 321 - 9 .
35. Olson BM , Jankowska-Gan E , Becker JT , Vignali DA , Burlingham WJ , McNeel DG . Human prostate tumor antigen-specific CD8+ regulatory T cells are inhibited by CTLA-4 or IL-35 blockade . J Immunol . 2012 ; 189 : 5590 - 01 .
36. Wu H , Li P , Shao N , Ma J , Ji M , Sun X , et al. Aberrant expression of Tregassociated cytokine IL-35 along with IL-10 and TGF-beta in acute myeloid leukemia . Oncol Lett . 2012 ; 3 : 1119 - 23 .
37. Zeng JC , Zhang Z , Li TY , Liang YF , Wang HM , Bao JJ , et al. Assessing the role of IL-35 in colorectal cancer progression and prognosis . Int J Clin Exp Pathol . 2013 ;6: 1806 - 16 .
38. Jin P , Ren H , Sun W , Xin W , Zhang H , Hao J. CirculatingIL -35 in pancreatic ductal adenocarcinoma patients . Hum Immunol . 2014 ; 75 : 29 - 33 .
39. Lin Y , Huang Y , Lu Z , Luo C , Shi Y , Zeng Q , et al. Decreased plasma IL-35 levels are related to the left ventricular ejection fraction in coronary artery diseases . PLoS One . 2012 ; 7 : e52490 .
40. Mao H , Gao W , Ma C , Sun J , Liu J , Shao Q , et al. Human placental trophoblasts express the immunosuppressive cytokine IL-35 . Hum Immunol . 2013 ; 74 : 872 - 7 .
41. Chen C , Deng Y , Chen H , Wu X , Cheng S , Xu Y , et al. Decreased concentration of IL-35 in plasma of patients with asthma and COPD . Asian Pac J Allergy Immunol . 2014 ; 32 : 211 - 7 .
42. Cooper AM , Kipnis A , Turner J , Magram J , Ferrante J , Orme IM . Mice lacking bioactive IL-12 can generate protective, antigen-specific cellular responses to mycobacterial infection only if the IL-12 p40 subunit is present . J Immunol . 2002 ; 168 : 1322 - 7 .
43. Conti HR , Shen F , Nayyar N , Stocum E , Sun JN , Lindemann MJ , et al. Th17 cells and IL-17 receptor signaling are essential for mucosal host defense against oral candidiasis . J Exp Med . 2009 ; 206 : 299 - 11 .
44. Shi YY , Dai MJ , Wu GP , Zhou PP , Fang Y , Yan XB . Levels of interleukin-35 and its relationship with regulatory T-cells in chronic hepatitis B patients . Viral Immunol . 2015 ; 28 : 93 - 100 .