Th1-Th17 Cells Contribute to the Development of CP/CPPS

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Wolfcub
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Th1-Th17 Cells Contribute to the Development of CP/CPPS

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PLoS One. 2013; 8(4): e60987.
Published online 2013 April 5. doi: 10.1371/journal.pone.0060987PMCID: PMC3618515


Th1-Th17 Cells Contribute to the Development of Uropathogenic Escherichia coli-Induced Chronic Pelvic Pain
Marsha L. Quick,#1 Larry Wong,#1 Soumi Mukherjee,1 Joseph D. Done,1 Anthony J. Schaeffer,1 and Praveen Thumbikat1,2,*
Jose Carlos Alves-Filho, Editor
1Department of Urology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States of America
2Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, United States of America
University of São Paulo, Brazil

Abstract
The etiology of chronic prostatitis/chronic pelvic pain syndrome in men is unknown but may involve microbes and autoimmune mechanisms. We developed an infection model of chronic pelvic pain in NOD/ShiLtJ (NOD) mice with a clinical Escherichia coli isolate (CP-1) from a patient with chronic pelvic pain. We investigated pain mechanisms in NOD mice and compared it to C57BL/6 (B6) mice, a strain resistant to CP-1-induced pain. Adoptive transfer of CD4+ T cells, but not serum, from CP-1-infected NOD mice was sufficient to induce chronic pelvic pain. CD4+ T cells in CP-1-infected NOD mice expressed IFN-γ and IL-17A but not IL-4, consistent with a Th1/Th17 immune signature. Adoptive transfer of ex-vivo expanded IFN-γ or IL-17A-expressing cells was sufficient to induce pelvic pain in naïve NOD recipients. Pelvic pain was not abolished in NOD-IFN-γ-KO mice but was associated with an enhanced IL-17A immune response to CP1 infection. These findings demonstrate a novel role for Th1 and Th17-mediated adaptive immune mechanisms in chronic pelvic pain.
The Discussion Section:
Discussion
CP/CPPS has been postulated to be associated with microbial insult to the prostate but the evidence for this etiology has been hard to pinpoint with disease diagnosis occurring at a time-point discrete from the inciting event. In an earlier study, we presented the first experimental evidence that a bacterial isolate from a patient with CP/CPPS can initiate and sustain the development of chronic pelvic pain, a distinguishing characteristic of CP/CPPS [8]. However, beyond demonstration of an important role for the host genetic background and the virulence of the infecting microbe, the mechanism underlying chronic pelvic pain remained to be solved. In this study we demonstrate that chronic pelvic pain initiated though a microbial infection of the prostate is induced by a Th1/Th17 immune response that is closely tied to the nature of the offending pathogen and the genetic background of the infected host.

Studies from a number of laboratories, including ours, have demonstrated the ability of UPEC delivered transurethrally to infect the murine prostate and induce inflammation [8], [13]–[16]. In our previous study utilizing the CP1 bacterial isolate, the presence of inflammation five days after infection was observed in NOD and B6 mice and not observed to correlate with the development of chronic pelvic pain. Consistent with these earlier findings, chronic inflammation at 28 days, as measured by leukocytic infiltration, did not appear to be different between the two strains of mice (Fig. 1A). Examination of the chemokine profiles induced in the prostate revealed differential expression of a small number of cytokines/chemokines in NOD and B6 mice (Fig. 1D). In NOD mice, elevation of the CXC chemokine MIP-2 is suggestive of neutrophil activation and recruitment [17]. In B6 mice, elevated expression of IL-16, a pro-inflammatory cytokine chemotactic for CD4+ T lymphocytes, monocytes, and eosinophils and IL-1ra, a receptor antagonist of IL-1 cytokines was observed. The increased size of secondary lymphoid organs in the NOD mouse is suggestive of robust immune activation in NOD mice.

Chronic prostatitis in humans is associated with the presence of specific auto-antibodies [18] as well as auto-reactive T cells [19]–[22]. In this study, we demonstrate that adoptive transfer of T cells and more specifically CD4+ T cells from CP-1 infected donors is sufficient to mediate the development of pelvic pain. Our results are consistent with a reported requirement for CD4+ T cells in the NOD autoimmune prostatitis model [23] as well as a well-studied requirement for CD4+ T cells in the pathogenesis of type-1 diabetes in the NOD genetic background [24]. CD8+ T cells, however, displayed an ability to mediate pelvic pain irrespective of whether they were derived from naïve or CP1-infected NOD mice. These nonspecific results suggest that endogenous, antigen-specific CD8+ T cells in naïve NOD mice are capable of undergoing activation without co-stimulation [25], [26] and expanding in an non-specific antigen-independent manner [27] to mediate pelvic pain. We have previously demonstrated that infection of the NOD prostate by CP1 or the closely related UPEC cystitis isolate NU14 lead to different pelvic pain outcomes, with CP1 alone producing sustained chronic pelvic pain [8]. In this study, we show that lymph node derived CD4+ T cells from CP1 infection are characterized by significant intracellular expression of IFN-γ, a signature cytokine for Th1 helper T cells. NU14 by contrast was incapable of such a response indicating the pathogen specificity of the immune response. The absence of IL-4, signifying a lack of a Th2 response to CP1 is consistent with the inability of immune serum to transfer chronic pelvic pain. Our results agree with previous studies demonstrating an important role for IFN-γ in the development of autoimmune prostatitis in the NOD mouse [28], but represents, to the best of our knowledge, the first demonstration of a Th1 response that leads to chronic pelvic pain. In contrast to the immune response in the local lymph node, infiltrating CD4+ T lymphocytes in the NOD prostate were characterized by increased IFN-γ and IL-17A expression in response to CP1 infection. IL-17A has been reported to be elicited from γδ T cells in UPEC bladder infections in a murine model and appear to be primarily functional at the level of innate immunity [29]. In contrast, our results demonstrate expression of IL-17A in αβ CD4+ T cells in a host and pathogen specific manner. Expectedly, the increase in IFN-γ and IL-17A transcription factors is associated with a diminished regulatory T cell response.

Beyond mere association with chronic pain responses, both IL-17A and IFN-γ appear to be sufficient to mediate chronic pelvic pain. IL-17A is a potent regulator of matrix metalloproteases, acute phase proteins, IL-6, and CXC chemokines that function to recruit neutrophils leading to their mobilization, recruitment, and activation. IL-17A may mediate hyperalgesia and pronociceptive effects through neutrophils [30] or alternatively through direct mechanisms that contribute to sensitization and allodynia [31]–[33]. In this context, it is worth noting that IL-17A has been demonstrated to be an upstream mediator of CCL2/MCP-1 expression [34]. CCL2/MCP-1 is elevated in prostate secretions of men with CP/CPPS and is a potential biomarker with a role in CP/CPPS pathogenesis [35]. IFN-γ, a well-known pro-inflammatory cytokine has also been implicated in hypernociception [36] and neuropathic hypersensitivity through T cell activation induced mechanisms [37]. Our studies show an exacerbated pain response in NOD-IFN-γ-KO mice when compared to NOD mice infected with CP1. Furthermore, while IL-17 was not expressed in the draining lymph nodes of wild type NOD mice, IL-17 is greatly expressed in the lymph nodes of IFN-γ-KO CP-1-infected mice. These observations while counterintuitive, are supported by studies in autoimmune disease models such as experimental autoimmune encephalitis (EAE) where, in the absence of IFN-γ, disease progression in EAE is exacerbated and IFN-γ is believed to act as an inhibitor of disease initiation [38]–[41]. The key cytokine responsible for the proinflammatory autoimmunity in these models appear to be IL-17A [38] and IFN-γ is known to inhibit Th17 polarization. Thus, in its absence, Th17 development and IL-17A expression is expected to progress unimpeded and promote disease/inflammation. Taken together, it is not surprising that the increase in pelvic pain observed in the NOD-IFN-γ-KO mice is associated with an increase in IL-17A.

In summary, we show that a Th1 and Th17 immune response is responsible for pain in CP-1-infected NOD mice while C57BL/6 mice infected with CP-1 and NOD mice infected with NU14 fail to mount a Th1/Th17 immune response. Our finding that antigen-experienced Th1 and Th17 cells transfer pain into naïve mice demonstrates that inflammatory T cells are the main mediators of pain in the mouse infection model of CP/CPPS. Important questions that remain to be answered and a future focus of our studies is identifying the effector cells/mechanisms activated by IL-17A and IFN-γ in the prostate that mediate disease pathology and symptoms. In the context of human CP/CPPS, our results strongly argue for systematic examination of the immune profile of patients with chronic pain to identify subsets of individuals that may have active Th1/Th17 mechanisms. Thus, these results have potentially important implications in understanding the pathogenesis of CP/CPPS.
Age: 30 | Onset Age: 19| Symptoms: Urinary frequency, Urinary urgency, constant 24/7 sensation in the penis (in the tip mainly - burning/pressure/discomfort/"wetness"), Nocturia, discomfort and pressure in the pelvic region radiating to the abdomen and becoming severe as time passes since last urination (resolved in 2014 by myofascial release), Stream velocity is somewhat slow and prolonged with an average velocity of ~13cc/min (and max 18cc/min) found in flowmetry test when bladder is filled with 500cc at age 25 (I always feel like I need to press my abdomen to urinate, improved later on when using alpha blockers)| Helped By: especially MYOFASCIAL RELEASE (especially in the areas of hips and abdomen) - generally resolved my abdominal aches, but penile symptoms remained the same| Worsened By: Coffee and possibly some other food as well| Other comments: Quercetin and acupuncture helped me no more than a placebo effect. Age 25-26: Diagnosed with indirect inguinal hernia and medium hydrocele at the same side. After operation many of the acute symptoms disappeared, but the chronic urinary and pelvic symptoms remained much the same.
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Re: Th1-Th17 Cells Contribute to the Development of CP/CPPS

Post by webslave »

Full article available here
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3618515/

We've long known that an infection can induce chronic pelvic pain. This study attempts to determine how that can happen. Note that there were no bacteria in the prostates of the mice after 30 days:
We also examined bacterial colonization of the prostates of these mice at day 30 with CP1 infection and did not observe any significant bacterial colonization
So pain —determined in mice by monitoring "tactile allodynia" (pain elicited by touching the mice)— persisted in a particular strain of mice (non-obese diabetic or "NOD" mice, which exhibit a susceptibility to spontaneous development of autoimmune disease) long after the infection passed.

Note also that the E. coli bacterial isolate from a CP/CPPS patient, used to infect these mice, is in itself highly unusual. Very few CP/CPPS patients have E. coli in their prostates!

All that being said, the problem with murine (rodent) studies of a human pain syndrome is that rodents are not humans, and we don't know exactly what they are feeling, or whether their biochemical reactions are the same as ours in particular circumstances. The mouse with a floridly and severely inflamed prostate (autoimmune prostatitis), in obvious pain, is not the same as a human being with diffuse pelvic pain and an ostensibly normal prostate.

So studies such as this are interesting from a speculative viewpoint, but don't add much (or indeed anything) to the process of recovery for CP/CPPS patients.

This study argues for the testing of the immune system in CPPS patients, so you can expect more studies along these lines:
In the context of human CP/CPPS, our results strongly argue for systematic examination of the immune profile of patients with chronic pain to identify subsets of individuals that may have active Th1/Th17 mechanisms.
Th1 - http://en.wikipedia.org/wiki/Th1_cell
Th17 - http://en.wikipedia.org/wiki/T_helper_17_cell
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