Modeling and Experimental Analyses Reveals Signaling Plasticity in a Bi-Modular Assembly of CD40 Receptor Activated Kinases

Sarma, Uddipan; Sareen, Archana; Maiti, Manas Kumar; Kamat, Vanita; Sudan, Raki; Pahari, Sushmita; Srivastava, Neetu; Roy, Somenath; Sinha, Sitabhra; Ghosh, Indira; Chande, Ajit; Mukhopadhyaya, Robin; Saha, Bhaskar

doi:10.1371/journal.pone.0039898

Cited by 18 publications

(24 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Whereas such counteractive signaling pathways may be part of two antagonistic receptors such as CD28 and CD152, we established the single receptor plasticity whereby CD40 signals reciprocally through ERK-1/2 and p38MAPK that result in counteractive functions such as parasite survival or parasite elimination, respectively (5). We established a novel bimodular architecture of the CD40 signaling pathway (23) and showed that L. major switches CD40 signaling from the p38MAPK module to the ERK-1/2 module. Because of two transmodular feedback loops, activation of ERK-1/2 in one module reduces p38MAPK activation in the other module.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of CD40-Induced N-Ras Activation Reduces Leishmania major Infection

Chakraborty

Srivastava

Jha

et al. 2015

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

Leishmania major is a parasite that resides and replicates in macrophages. We previously showed that the parasite enhanced CD40-induced Raf-MEK-ERK signaling but inhibited PI3K-MKK-p38MAPK signaling to proleishmanial effects. As Raf and PI3K have a Ras-binding domain but exert opposite effects on Leishmania infection, we examined whether Ras isoforms had differential roles in Leishmania infection. We observed that L. major enhanced N-Ras and H-Ras expression but inhibited K-Ras expression in macrophages. L. major infection enhanced N-Ras activity but inhibited H-Ras and K-Ras activity. TLR2 short hairpin RNA or anti-TLR2 or anti-lipophosphoglycan Abs reversed the L. major–altered N-Ras and K-Ras expressions. Pam3CSK4, a TLR2 ligand, enhanced N-Ras expression but reduced K-Ras expression, indicating TLR2-regulated Ras expression in L. major infection. Whereas N-Ras silencing reduced L. major infection, K-Ras and H-Ras silencing enhanced the infection both in macrophages in vitro and in C57BL/6 mice. BALB/c-derived macrophages transduced with lentivirally expressed N-Ras short hairpin RNA and pulsed with L. major–expressed MAPK10 enhanced MAPK10-specific Th1-type response. CD40-deficient mice primed with these macrophages had reduced L. major infection, accompanied by higher IFN-γ but less IL-4 production. As N-Ras is activated by Sos, a guanine nucleotide exchange factor, we modeled the N-Ras–Sos interaction and designed two peptides from their interface. Both the cell-permeable peptides reduced L. major infection in BALB/c mice but not in CD40-deficient mice. These data reveal the L. major–enhanced CD40-induced N-Ras activation as a novel immune evasion strategy and the potential for Ras isoform–targeted antileishmanial immunotherapy and immunoprophylaxis.

show abstract

Section: Discussionmentioning

confidence: 99%

Inhibition of CD40-Induced N-Ras Activation Reduces Leishmania major Infection

Chakraborty

Srivastava

Jha

et al. 2015

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

“…The quality of the response pertains to its counteractively functioning elements. Using CD40 as a model receptor with crucial roles in diverse immune functions, we previously showed that such counteractive functions or the quality of the response-proinflammatory or antiinflammatory-are regulated by a signaling system with signal strength calibrating feedback loops (7,10). The CD40 signaling system consists of cascades of kinases in two modules: one module transmits CD40 signal through lyn and p38MAPK while the other module transmits CD40 signal through syk and ERK-1/2.…”

Section: Discussionmentioning

confidence: 99%

“…In macrophages, a strong CD40 stimulation induces preferential phosphorylation of p38MAPK and proinflammatory cytokine IL-12 production, whereas a weaker CD40 stimulation induces ERK-1/2 phosphorylation and anti-inflammatory cytokine IL-10 production (5,6). This CD40 signaling reciprocity emerges because of sustained p38MAPK and ERK-1/2 counterregulation effected by two trans-modular feedback loops (7). This emergent property of the CD40 signaling pathway implies that the threshold for activation of p38MAPK can be calibrated by regulating these MAPK-oriented feedback loops (8).…”

mentioning

confidence: 99%

SHP-1 Plays a Crucial Role in CD40 Signaling Reciprocity

Khan

Srivastava

et al. 2014

The Journal of Immunology

Self Cite

View full text Add to dashboard Cite

CD40 plays dual immunoregulatory roles in Leishmania major infection and tumor regression. The functional duality emerges from CD40-induced reciprocal p38MAPK and ERK-1/2 phosphorylations. Because phosphotyrosine-based signaling in hematopoietic cells is regulated by the phosphotyrosine phosphatase SHP-1, which is not implied in CD40 signaling, we examined whether SHP-1 played any roles in CD40-induced reciprocal signaling and anti-leishmanial function. We observed that a weaker CD40 stimulation increased SHP-1 activation. ERK-1/2 inhibition or p38MAPK overexpression inhibited CD40-induced SHP-1 activation. An ultra-low-dose, CD40-induced p38MAPK phosphorylation was enhanced by SHP-1 inhibition but reduced by SHP-1 overexpression. A reverse profile was observed with ERK-1/2 phosphorylation. SHP-1 inhibition reduced syk phosphorylation but increased lyn phosphorylation; syk inhibition reduced but lyn inhibition enhanced CD40-induced SHP-1 phosphorylation. Corroborating these findings, in L. major–infected macrophages, CD40-induced SHP-1 phosphorylation increased and SHP-1 inhibition enhanced CD40-induced p38MAPK activation and inducible NO synthase expression. IL-10 enhanced SHP-1 phosphorylation and CD40-induced ERK-1/2 phosphorylation but reduced the CD40-induced p38MAPK phosphorylation, whereas anti–IL-10 Ab exhibited reverse effects on these CD40-induced functions, identifying IL-10 as a crucial element in the SHP-1-MAPK feedback system. Lentivirally overexpressed SHP-1 rendered resistant C57BL/6 mice susceptible to the infection. Lentivirally expressed SHP-1 short hairpin RNA enhanced the CD40-induced L. major parasite killing in susceptible BALB/c mice. Thus, we establish an SHP-1–centered feedback system wherein SHP-1 modulates CD40-induced p38MAPK activation threshold and reciprocal ERK-1/2 activation, establishing itself as a critical regulator of CD40 signaling reciprocity and mechanistically re-emphasizing its role as a potential target against the diseases where CD40 is involved.

show abstract

“…CD40 signaling pathway is characterized by bimodular architecture which endows the system to respond to CD40 engagement depending on CD40 surface expression and consequently the strength of the signal dose, thus reciprocally controlling ERKI/2 and p38 MAPK activation and eventually IL-12 or IL-I 0 production (27). In the case of DCs, CD40 engagement stimulates the activation of The phosphorylation of these proteins usually leads to production of IL-12 and up-regulation of co-stimulatory molecule expression (28).…”

Section: Inhib Ition Oferki/2 Activation Up-regulates Il-1 2/ Il-io Rmentioning

confidence: 99%

Low CD40 Expression Levels in Leishmania Infantum-Infected Bone Marrow Dendritic Cells Evoke Regulatory Responses by Down-Regulating Interleukin-12 Production: Role of ERK1/2

2014

View full text Add to dashboard Cite

Dendritic cells (DCs) playa pivotal role in promoting resistance to leishmaniasis, both by activating CD4+ T cells and endorsing their differentiation into Th 1 cells by producing interleukin (IL)-12. High level ofIL-12 production, a decisive component ofthe DC maturation, requires not only microbial stimuli but also strong CD40-CD40L interactions. Until now, the mechanisms by which Leishmania (L.) infantum parasites affect DC functional maturation and consequently T cell polarization are not fully understood.In the present study, we investigated the response that is elicited when L. infantum promastigoteinfected bone marrow-derived DCs (BM-DCs) to CD40 engagement and this way mimicking DC-T cells interactions at the early stages of infection. We found that L. infantum promastigotes-infected BM-DCs following CD40 engagement were capable of inducing significant amounts ofTNF-a and IL-IO, whereas IL-12 production remained unaffected compared to infected untreated cells. Interestingly, infected BM-DCs did not up-regulate CD40 surface expression. On the other hand, BM-DC stimulation with soluble Leishmania antigen (SLA) resulted not only in significant increase of co-stimulatory molecule expression but also IL-12 and IL-IO production. CD40 engagement on L. infantum-infected BM-DCs sustained ERK1I2 activation induced by the parasite alone. Inhibition of ERK1I2 activation with the use of PD98059 inhibitor prior to CD40 engagement on L. infantum-infected BM-DCs resulted in significant up-regulation ofp38 MAPK phosphorylation and IL-12 production, whereas it did not affect TNF-a and IL-IO production. These findings suggest that L. infantum has evolved specific strategies to avoid efficient DC-T cell interactions by suppressing CD40 expression and consequently leading CD40 signaling pathways to ERK1I2 activation.

show abstract

Modeling and Experimental Analyses Reveals Signaling Plasticity in a Bi-Modular Assembly of CD40 Receptor Activated Kinases

Cited by 18 publications

References 25 publications

Inhibition of CD40-Induced N-Ras Activation Reduces Leishmania major Infection

Inhibition of CD40-Induced N-Ras Activation Reduces Leishmania major Infection

SHP-1 Plays a Crucial Role in CD40 Signaling Reciprocity

Low CD40 Expression Levels in Leishmania Infantum-Infected Bone Marrow Dendritic Cells Evoke Regulatory Responses by Down-Regulating Interleukin-12 Production: Role of ERK1/2

Contact Info

Product

Resources

About