Tumor-associated immune cell infiltrate density in penile squamous cell carcinomas

Penile squamous cell carcinomas are rare tumor entities throughout Europe. Early lymphonodal spread urges for aggressive therapeutic approaches in advanced tumor stages. Therefore, understanding tumor biology and its microenvironment and correlation with known survival data is of substantial interest in order to establish treatment strategies adapted to the individual patient. Fifty-five therapy naïve squamous cell carcinomas, age range between 41 and 85 years with known clinicopathological data, were investigated with the use of tissue microarrays (TMA) regarding the tumor-associated immune cell infiltrate density (ICID). Slides were stained with antibodies against CD3, CD8 and CD20. An image analysis software was applied for evaluation. Data were correlated with clinicopathological characteristics and overall survival. There was a significant increase of ICID in squamous cell carcinomas of the penis in relation to tumor adjacent physiological tissue. Higher CD3-positive ICID was significantly associated with lower tumor stage in our cohort. The ICID was not associated with overall survival. Our data sharpens the view on tumor-associated immune cell infiltrate in penile squamous cell carcinomas with an unbiased digital and automated cell count. Further investigations on the immune cell infiltrate and its prognostic and possible therapeutic impact are needed.

Pathogenetic value of cell infiltrate in immunoinflammatory rheumatic diseases

Cell infiltrate is a morphological substrate of immunoinflammatory rheumatic diseases. The systemic wide progressive disorganization of loose fibrous connective tissue is accompanied by the loss of tolerance with its own autoantigenes, activation of macrophagal-monocyte cells and autoreactive clones of T and B lymphocytes. Hyperproduction of pro-inflammatory chemokines and cytokines, local adhesive ligandreceptor interactions, endothelial reaction and angiogenesis contribute to the formation of cell infiltrate, ectopic lymphoid structures and GZT-granulomas in situ. The autoimmune response is the result of successive systemic and local molecular cellular events in which the mechanisms of congenital and adaptive immunity are involved. When interpreting immunopathogenesis of rheumatic diseases, all models and schemes adopted in the field of fundamental immunology are used. This is a model of MHC-restrictions, a model of molecular mimicry, or cross of the antigen presentation, a model of disrupting central or peripheral tolerance to auto-antigens, a model of candidate “triggers” of autoimmune and autoinflammatory processes, a model of associations of alleles MHC I and II classes with specific, nosologically unique, rheumatic diseases.

The effect of clozapine and MIS416 on demyelination, remyelination and immunomodulation in the murine cuprizone model

<p>Multiple Sclerosis (MS) is a disorder of the central nervous system that affects approximately 2.5 million people worldwide. Due to the heterogeneous nature of the disease, and the want of an identified cause, treatment of MS remains difficult. Treatments are available, however these are limited in efficacy and are not suitable for all forms of MS. Disease pathology is characterised by the formation of demyelinating lesions in the central nervous system (CNS) which lead to cognitive and motor impairments associated with the disease. These CNS lesions can be classified as those with immune cell involvement or those without immune cell infiltrate, which are more commonly seen in progressive forms of MS, and currently, there are no treatments available for progressive MS. Due to the limited options available for treating progressive MS, this thesis aims to identify the therapeutic effect provided by the immunomodulatory compounds, MIS416 and clozapine, in a non-immune mediated model of MS, which is believed to more closely resemble progressive disease. Both of these compounds have been shown previously to reduce disease burden in an immune-driven animal model of MS. To investigate the effect of immune-modulating therapies on lesions without immune cell infiltrate, the cuprizone model of non-immune demyelination was used. In summary, the work presented in this thesis found that treatment with MIS416 and clozapine led to improved performance on behavioural assays, although neither agent inhibited cuprizone-induced demyelination or enhanced remyelination. The cellular mechanism behind the observed behavioural improvement is yet to be confirmed. MIS416 was able to maintain its previously identified immunomodulatory properties when administered in this novel setting. Moreover, novel changes to serum growth factors were identified that could provide unexpected benefit to MS patients administered MIS416. In addition to reversing cuprizone-induced behavioural deficits, clozapine reduced LPS-driven inflammatory cytokine production by microglia, indicating that clozapine has the ability to directly reduce inflammation, which may benefit progressive MS patients. Protective effects provided by either of these compounds could aid in the development of unique combination therapies to target both the inflammatory immune component and the cellular components seen at different stages of MS. MIS416 induced changes to serum cytokines and growth factors in the periphery could be harnessed to treat not just MS but other auto-immune diseases characterised by a similar cytokine profile.</p>

The effect of clozapine and MIS416 on demyelination, remyelination and immunomodulation in the murine cuprizone model

<p>Multiple Sclerosis (MS) is a disorder of the central nervous system that affects approximately 2.5 million people worldwide. Due to the heterogeneous nature of the disease, and the want of an identified cause, treatment of MS remains difficult. Treatments are available, however these are limited in efficacy and are not suitable for all forms of MS. Disease pathology is characterised by the formation of demyelinating lesions in the central nervous system (CNS) which lead to cognitive and motor impairments associated with the disease. These CNS lesions can be classified as those with immune cell involvement or those without immune cell infiltrate, which are more commonly seen in progressive forms of MS, and currently, there are no treatments available for progressive MS. Due to the limited options available for treating progressive MS, this thesis aims to identify the therapeutic effect provided by the immunomodulatory compounds, MIS416 and clozapine, in a non-immune mediated model of MS, which is believed to more closely resemble progressive disease. Both of these compounds have been shown previously to reduce disease burden in an immune-driven animal model of MS. To investigate the effect of immune-modulating therapies on lesions without immune cell infiltrate, the cuprizone model of non-immune demyelination was used. In summary, the work presented in this thesis found that treatment with MIS416 and clozapine led to improved performance on behavioural assays, although neither agent inhibited cuprizone-induced demyelination or enhanced remyelination. The cellular mechanism behind the observed behavioural improvement is yet to be confirmed. MIS416 was able to maintain its previously identified immunomodulatory properties when administered in this novel setting. Moreover, novel changes to serum growth factors were identified that could provide unexpected benefit to MS patients administered MIS416. In addition to reversing cuprizone-induced behavioural deficits, clozapine reduced LPS-driven inflammatory cytokine production by microglia, indicating that clozapine has the ability to directly reduce inflammation, which may benefit progressive MS patients. Protective effects provided by either of these compounds could aid in the development of unique combination therapies to target both the inflammatory immune component and the cellular components seen at different stages of MS. MIS416 induced changes to serum cytokines and growth factors in the periphery could be harnessed to treat not just MS but other auto-immune diseases characterised by a similar cytokine profile.</p>

Therapeutic Role of Platelet Rich Plasma in Formaldehyde-Induced Arthritis in Adult Male Albino Rats

Introduction Osteoarthritis (OA) is a common health problem. Platelet-rich plasma (PRP) has been recognized to enhance articular cartilage metabolism. Aim of the work the study was designed to investigate the influence of PRP on cartilage healing after induction of arthritis. Material and methods Forty two adult male albino rats were used in this study. The rats were randomly divided into three groups: Group I (n = 18): the control group (Ia, Ib & Ic) Ib & Ic were injected intra-articularly with saline and left for 3 and 6 weeks. Group II (n = 12): arthritic group, in which osteoarthritis was induced by injection of 0.02ml 5% formaldehyde once in the right knee joints, left without treatment, and were sacrificed after three weeks (IIa) or after six weeks (IIb). Group III(n = 12): arthritis was induced as group II, one week later, the rats were intra-articularly injected with single dose of 0.3ml PRP in the same joint then were sacrificed three weeks (IIIa) or six weeks (IIIb) after formaldehyde injection. At the end of the study the right knee joints were taken, decalcified then processed for paraffin sections to be examined by light microscope using H&E, toluidine blue and Masson’s trichrome (MTC) stains. Immunohistochemistry for caspase-3 enzyme was done to demonstrate apoptotic chondrocytes. Morphometric study was conducted to measure the thickness of the non-calcified cartilage, count the chondrocytes and synovial membrane inflammatory cells and Mankin's score. Then statistical analysis was done. Results The arthritic groups revealed irregular surface of the articular cartilage, loss of the articular matrix and bone eburnation. Moreover, there was apparent hypocellularity and disorganization of the chondrocytes. Osteoclasts and osteoblasts were seen invading the osteochondral junction. MTC stained sections of the synovial membrane showed deposition of thick collagen bundles with heavy inflammatory cell infiltrate and numerous blood vessels. The affinity of the articular cartilage to toluidine blue stain was apparently decreased while caspase-3 immunoreactivity was apparent in many chondrocytes .Group IIIa demonstrated almost similar histological findings as the control group; regular articular cartilage surface with regularly arranged chondrocytes in the different cartilage zones. Synovial membrane illustrated minimal inflammatory cell infiltrate with thin collagen bundles and small blood vessels in MTC stained sections. There was high affinity of the articular cartilage to the toluidine blue stain and few chondrocytes showed positive caspase-3 immunoreactivity. Group IIIb revealed continuous surface of the articular cartilage, yet with minimal fibrillation in some areas. Osteoblasts and osteoclasts were seen invading the calcified cartilage. Synovial membrane showed deposition of dense collagen bundles with some inflammatory cell infiltrate. Toluidine blue sections revealed decreased articular cartilage affinity to the stain while caspase-3 immunoreactivity was evident in many chondrocytes. The morphometric results and statistical analysis confirmed the histological findings. Conclusion Intra-articular injection of PRP demonstrated advantageous role on articular cartilage healing, however, these effects appeared to be transient. So the need of multiple injections of PRP has to be considered in cases of OA.

TCR Clonality and Genomic Instability Signatures as Prognostic Biomarkers in High Grade Serous Ovarian Cancer

Purpose: Immune infiltration is a prognostic factor in high-grade serous ovarian carcinoma (HGSC) but immunotherapy efficacy is disappointing. Genomic instability is now used to guide the therapeutic value of PARP inhibitors. We aimed to investigate exome-derived parameters to assess the tumor microenvironment according to genomic instability profile. Methods: We used the HGSC TCGA (the cancer genome atlas) dataset with genomic characteristics, including homologous recombination deficiency (HRD), copy number variant (CNV) signatures, TCR (T cell receptor) clonality and abundance of tissue-infiltrating immune and stromal cell populations. We then investigated the relationship with survival data. Results: In 578 HGSC patients, HRD status, CNV signature 7 and TCR clonality were associated with longer survival. The combination of high CNV signature 7 expression and HRD status or high CNV signature 3 expression and high TCR clonality was associated with a trend towards longer survival compared to each variable alone. Combining T cell infiltrate and TCR clonality improved the prognostic value compared to T cells infiltration alone. Prognostic value of TCR clonality was confirmed in an independent cohort. Conclusions: TCR clonality is an emerging prognostic biomarker that improves T cell infiltrate information. Analysis of TCR clonality combined with genomic instability could be an interesting prognostic biomarker.

The peri- and intratumoral immune cell infiltrate and PD-L1 status in invasive squamous cell carcinomas of the penis

Introduction Penile carcinomas are rare tumors throughout Europe. Therefore, little attention is drawn to this disease. That makes it important to study tumor-associated key metrics and relate these to known data on penile neoplasias. Materials and methods A cohort of 60 well-defined penile invasive carcinomas with known human papillomavirus (HPV) infection status was investigated. Data on tumor type, grading and staging were recorded. Additionally, data on the peri- and intratumoral immune cell infiltrate in a semiquanititave manner applying an HE stain were assessed. Results Our study showed a significant correlation of immune cell infiltrate and pT stage with overall survival. Therefore, in a subset of tumors, PD-L1 staining was applied. For tumor proportion score (TPS), 26 of 30 samples (87%) were scored >0%. For the immune cell score (IC), 28 of 30 samples (93%) were defined as >0% and for CPS, 29 of 30 samples (97%) scored >0. PD-L1 expression was not associated with overall survival. Conclusion PD-L1 is expressed in penile carcinomas, providing a rationale for targeted therapy with checkpoint inhibitors. We were able to show that immune reaction appears to be prognostically relevant. These data enhance the need for further studies on the immune cell infiltrate in penile neoplasias and show that PD-L1 expression is existent in our cohort, which may be a potential target for checkpoint inhibitor therapy.

Immune Checkpoint Inhibitors Mediated Lymphocytic and Giant Cell Myocarditis: Uncovering Etiological Mechanisms

The advent of immune checkpoint inhibitors (ICIs) has revolutionized the field of oncology, but these are associated with immune related adverse events. One such adverse event, is myocarditis, which has limited the continued immunosuppressive treatment options in patients afflicted by the disease. Pre-clinical and clinical data have found that specific ICI targets and precipitate distinct myocardial infiltrates, consistent with lymphocytic or giant cell myocarditis. Specifically, it has been reported that CTLA-4 inhibition preferentially results in giant cell myocarditis with a predominately CD4+ T cell infiltrate and PD-1 inhibition leads to lymphocytic myocarditis, with a predominately CD8+ T cell infiltrate. Our manuscript discusses the latest literature surrounding ICI pathways and targets, while detailing proposed mechanisms behind ICI mediated myocarditis.