Intraepithelial Infiltration Research Articles

Intraepithelial CD15 infiltration identifies high-grade anal dysplasia in people with HIV.

Men who have sex with men (MSM) with HIV are at high risk for squamous intraepithelial lesion (SIL) and anal cancer. Identifying local immunological mechanisms involved in the development of anal dysplasia could aid treatment and diagnostics. Here, we studied 111 anal biopsies obtained from 101 MSM with HIV, who participated in an anal screening program. We first assessed multiple immune subsets by flow cytometry, in addition to histological examination, in a discovery cohort. Selected molecules were further evaluated by immunohistochemistry in a validation cohort. Pathological samples were characterized by the presence of resident memory T cells with low expression of CD103 and by changes in natural killer cell subsets, affecting residency and activation. Furthermore, potentially immunosuppressive subsets, including CD15+CD16+ mature neutrophils, gradually increased as the anal lesion progressed. Immunohistochemistry verified the association between the presence of CD15 in the epithelium and SIL diagnosis for the correlation with high-grade SIL. A complex immunological environment with imbalanced proportions of resident effectors and immune-suppressive subsets characterized pathological samples. Neutrophil infiltration, determined by CD15 staining, may represent a valuable pathological marker associated with the grade of dysplasia.

Abstract 169: Prevalence, immune checkpoint expression and spatial interplay of intraepithelial lymphocytes, macrophages, and dendritic cells in 4740 human carcinomas

Abstract Background: Although there is raising evidence that the density and the level of immune checkpoint expression of immune cell subpopulations that are in direct contact to the tumor cells (intraepithelial) can predict response to immune checkpoint therapy and patient’s outcome, a comprehensive assessment of intraepithelial immune cell subpopulations and their spatial interplay is lacking. Material and methods: To comprehensively assess the density, level of immune checkpoint expression, and spatial interplay of 48 immune checkpoint expressing intraepithelial leukocyte subpopulations in 46 carcinoma entities, 4740 tumor samples in a tissue microarray format were stained with 21 antibodies using our BLEACH&STAIN mfIHC approach. A deep learning-based framework comprising 18 different convolutional neuronal networks (U-Net and DeepLabv3+) were used for image analysis. Results: The mean intraepithelial immune cell density of CD8+ cytotoxic T-cells, CD4+ T-helper cells, FOXP3+ Tregs, CD20+ B-cells, M1/M2 macrophages and CD11c+ dendritic cells varied markedly between tumor entities and individual tumors. For example, 88 (±90) cells/mm2 were found in tubular breast cancer, 661 (±729) cells/mm2 in colorectal cancer, and 1280 (±1334) to 2325 (±2131) cells/mm2 in squamous cell cancers from various origins. The abundance of T-cell subpopulations, M2 macrophages, and dendritic cells was highly concordant (range r=0.49 to 0.92) across all tumor entities, while the density of M1 macrophages was unrelated (r=0.1, p<0.001). Unsupervised hierarchical clustering across all tumor entities revealed a cluster A of 634 patients from almost all different tumor entities with an exceptionally high density of intraepithelial immune cells, thus a highly inflamed tumor microenvironment that was characterized by the strongest interaction between intraepithelial CD8+ T-cell and CD4+ T-helper cells, M2 macrophages, as well as dendritic cells (DC, p<0.002 each) along with the highest expression level of TIM3, PD-1, CLTA-4 and PD-L1. In contrast, a cluster C of 2245 patients with the lowest intraepithelial immune cell infiltration was characterized by a significantly increased M1 proportion (p<0.001) and an enriched proportion of regulatory T-cells (p<0.001). Across all analyzed tumor entities, the intraepithelial highly inflamed cluster A was significantly linked to low pT (p<0.001). Conclusion: The data from this study provide a comprehensive characterization of intraepithelial immune cells across 46 different human carcinomas and identify an inflamed phenotype defined by strong interactions of intraepithelial CD8+ cytotoxic T-cells, CD4+ T-cells, dendritic cells, and M2 macrophages along with highest levels of TIM3, PD-1, PD-L1, and CTLA-4 expression that is linked to a favorable tumor phenotype. Citation Format: Zhihao Huang, Tim Mandelkow, Jan H. Müller, Andreas H. Marx, Till Krech, Guido Sauter, Julia Ebner, Maximilian Lennartz, Ronald Simon, Martina Kluth, Claudia Hube-Magg, Sarah Minner, Elena Bady, Niclas C. Blessin. Prevalence, immune checkpoint expression and spatial interplay of intraepithelial lymphocytes, macrophages, and dendritic cells in 4740 human carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 169.

Pathomorphological criteria and features of immune response in eosinophilic esophagitis and reflux esophagitis

Eosinophilic esophagitis (EoE) is an immune-mediated disease, manifested by dysphagia and characterized by intraepithelial infiltration: more than 15 eosinophils per field of view at x400 magnification, and requiring differential diagnosis with reflux esophagitis (RE). To access the implication of EoE histologic scoring system (EoEHSS) for differential diagnosis of EoE and RE and to characterize features of immune response in these diseases. 38 patients with EoE and 38 patients with RE were enrolled in the study. All the patients had esophagogastroduodenoscopy with biopsy. Biopsy specimens were stained with H&E and combined PAS/Alcian blue staining. Immunohistochemical evaluation was conducted with antibodies to CD3, CD4, CD8, CD20, CD56 and CD68. Grade score of EoEHSS in EoE was 2.4 times more than in RE (p<0.05). Stage score in EoE was 2.75 more than in RE (p<0.05). Intraepithelial count of CD3+ T-lymphocytes comprised 87 (76-95.5) in high-power view in EoE and 45 (38.5-48.5) in high-power view in RE. Intraepithelial count of CD4+ T-lymphocytes was 35 (28-41.5) in high-power view in EoE and 19 (16.5- 22.5) in high-power view in RE. Intraepithelial count of CD8+ T-lymphocytes comprised 59 (50.5-67.5) in high-power field in EoE and 27 (24-28.5) in high-power field in RE. The use of the EoEHSS histological rating scale for eosinophilic esophagitis is effective in the differential diagnosis of EoE and EC. Predominant cells in intraepithelial infiltrate are CD3+ T-lymphocytes both in EoE and RE, CD8+ cells prevail over CD4+ cells. In EoE intraepithelial count of CD3+ T-lymphocytes is 1.93 times more, count of intraepithelial CD4+ lymphocytes is 1.84 times more and count of CD8+ lymphocytes is 2.19 times more than in RE.

Эозинофильный эзофагит: что мы знаем и что мы можем?

Eosinophilic esophagitis is a chronic immune-mediated disease of the esophagus. It is characterized by severe eosinophilic inflammation of the esophageal mucosa and the develop-ment of submucosal fibrosis. Clinically, it is manifested by swallowing disorders. The pathogen-esis is based on genetically determined disorder of immune response and barrier function of the esophageal mucosa. The diagnosis of esophageal eosinophilic infiltration is established by clinical manifestations and morphology, after excluding diseases that cause esophageal eosinophilia. The criterion for diagnosis is intraepithelial eosinophilic infiltration with 15 or more eosinophils per field of view at magnification ×400 (or ≥60 eosinophils per 1 mm2). Secondary tests, such as measuring serum total immunoglobulin E levels, assessing peripheral blood eosinophilia, and skin allergy tests, are of lesser importance. The primary treatment for this condition is proton pump inhibitors. If the effect is insufficient, topical steroids and dietary restrictions may also be prescribed. Treatment effectiveness is evaluated 6–12 weeks after initiation by analyzing symp-toms and performing esophagogastroduodenoscopy with biopsy. Endoscopic dilatation is rec-ommended as a treatment option for patients experiencing severe dysphagia caused by esopha-geal narrowing. Biologic agents may also be considered as a possible treatment option. KEYWORDS: eosinophilic esophagitis, esophageal dysfunction, intraepithelial eosinophilic infiltration, proton pump inhibitors, topical steroids, diet, endoscopic dilatation, biologic agents. FOR CITATION: Belan E.B., Tibir'kova E.V. Eosinophilic esophagitis: current knowledge and management options? Russian Medical Inquiry. 2024;8(3):150–154 (in Russ.). DOI: 10.32364/2587-6821-2024-8-3-5.

Analysis of off-tumour toxicities of T-cell-engaging bispecific antibodies via donor-matched intestinal organoids and tumouroids

Predicting the toxicity of cancer immunotherapies preclinically is challenging because models of tumours and healthy organs do not typically fully recapitulate the expression of relevant human antigens. Here we show that patient-derived intestinal organoids and tumouroids supplemented with immune cells can be used to study the on-target off-tumour toxicities of T-cell-engaging bispecific antibodies (TCBs), and to capture clinical toxicities not predicted by conventional tissue-based models as well as inter-patient variabilities in TCB responses. We analysed the mechanisms of T-cell-mediated damage of neoplastic and donor-matched healthy epithelia at a single-cell resolution using multiplexed immunofluorescence. We found that TCBs that target the epithelial cell-adhesion molecule led to apoptosis in healthy organoids in accordance with clinical observations, and that apoptosis is associated with T-cell activation, cytokine release and intra-epithelial T-cell infiltration. Conversely, tumour organoids were more resistant to damage, probably owing to a reduced efficiency of T-cell infiltration within the epithelium. Patient-derived intestinal organoids can aid the study of immune–epithelial interactions as well as the preclinical and clinical development of cancer immunotherapies.

Eosinophil-mast cell pattern of intraepithelial infiltration as a marker of severity in CRSwNP

Chronic rhinosinusitis with nasal polyps (CRSwNP) is defined as a Type 2 eosinophilic disease, while CRSsNP is considered a Type 1 neutrophilic disease. Since neutrophils are also activated in eosinophilic CRSwNP, the eosinophil–neutrophil dualism has been revaluated. Among the inflammatory cells infiltrating sinus-nasal tissues, the role of mast cells (MCs) is not already recognized, although Clinical-Cytological Grading, which defines the severity of CRSwNP, attributes to mixed eosinophil-MC forms of CRSwNP a greater risk of recurrence. We aimed to examine nasal polyps from both a cytological and histopathological point of view, to evaluate the presence and localization of MCs. Cytological and histological examination of 39 samples of nasal polyps were performed. Immunohistochemistry was used to evaluate the presence of Tryptase + CD117 + MCs, which were counted both in the epithelial layer and in the lamina propria. A statistically significant correlation was found between intraepithelial MCs and CRSwNP severity (p < 0.001) and between the total eosinophil count and the total mast cell count (p < 0.001). Cytological examination and immunohistochemistry were comparable in detecting the presence of intraepithelial MCs (p = 0.002). The histological cut-off of 6 intraepithelial MCs was identified to detect severe CRSwNP (p < 0.001). MCs have been shown to be located in the lamina propria of almost all eosinophilic nasal polyps without significantly affecting their severity. Intraepithelial MCs are associated with greater severity of CRSwNP. Histopathological criteria of the eosinophil-MC form of CRSwNP in addition to the eosinophilic one, should be defined to guarantee patients effective and tailored treatments.

Immune features of ovarian tumors with a good response to neo-adjuvant chemotherapy in combination with an anti-PD-L1 alone or with an anti-CTLA4: Data from the randomized IneOV trial (a GINECO study).

5586 Background: We have shown that chemotherapy may have immunomodulatory properties and prime the tumor microenvironment (TME) by recruiting cytotoxic immune cells. We previously reported that neoadjuvant chemotherapy (NACT) associated with Durvalumab (D) +/- Tremelimumab (T) resulted in encouraging complete resection (70%) and complete pathological response (18%) rates in patients with unresectable ovarian cancer. Here, we aimed to 1) characterize the immune TME at baseline of responders compared to non-responders and 2) describe changes in immune TME in paired tumor samples at diagnosis and after 3 cycles of NACT with D +/- T in the randomized IneOV trial. Methods: Tumor samples from IneOV trial (55 at diagnosis, 40 after treatment) were analyzed for CD3+, CD68+ and CD20+ by multiplexed immunohistochemistry. Stromal and intra-epithelial TILs were scored as percentage of positive surface. Patients were classified as good pathological responders (pR) (CRS3) or pathological non-responders (pNR) (CRS1 or 2). Intra-tumoral T cell infiltration was described as High or Low (intraepithelial (ie)CD3+ &gt; median vs &lt; , respectively). Non parametric statistical tests were used. We evaluated the correlation of the markers with each others (Spearman test) and with pathological response rate (Mann-Withney test). Results: At diagnosis, in the intra-epithelial compartment of the TME, the most abundant cells were macrophages (median ieCD68+ = 3,4) compared to T (ieCD3+ = 1.1), B (ieCD20+ ND) ANOVA p-value = 0.002 and &lt; 0.0001 respectively . In individual tumors, infiltration by the 3 different immune subsets was poorly correlated suggesting immune TME heterogeneity in OC. Pathological response was evaluable in 64 patients: 19 pR and 45 pNR. At diagnosis, CD3+ (p = 0.02) and CD68+ (p = 0.006) infiltration was significantly higher in pR tumors compared to pNR. Most of pR (76%) had high intra-tumoral CD3+ infiltration versus 39% in pNR (p = 0.011). After treatment, tumors with pR showed significantly greater CD3+ intra-epithelial infiltration compared to pNR (p = 0.047). Conclusions: Our results suggest that good pathological response to NACT +D+/-T is associated with high levels of both T cells and macrophages in iTME at baseline, and increased intratumoral T cell infiltration post-treatment. Work is ongoing to further characterize the iTME, especially in pNR to identify other strategies to enhance the anti-tumor immune response. Clinical trial information: NCT03249142 .

Nasal cytology and histology in CRSwNP: Two sides of the same coin.

Knowledge of chronic rhinosinusitis with nasal polyps (CRSwNP) has increased rapidly over the past decade. However, the study of the histological features of nasal polyps has not gone hand in hand with the study of the inflammatory mechanisms underlying CRSwNP. Indeed, precisely because they are benign neoformations, nasal polyps have not attracted the attention of pathologists over the years. Nasal cytology has shown that CRSwNP, generally defined as a Type-2 disease, is characterized not only by eosinophilic but also mast cell inflammation and, in particular, the most severe forms of CRSwNP are precisely characterized by a mixed eosinophilic-mast cell inflammation. Interestingly, mast cells cannot be visualized by histology due to limitations in staining and magnification, and therefore are not commonly described in histological reports of nasal polyps. However, immunohistochemistry can highlight these latter cells and specifically this technique has recently demonstrated that mast cells are located in the lamina propria of almost all types of polyps and in the epithelial level of the most severe forms. Unfortunately, the latter technique is not commonly carried out in clinical practice by virtue of the high cost and time burden. On the other hand, nasal cytology is an easy-to-apply and economic diagnostic tool, commonly practiced in rhinological setting, which can effectively fill the gap between histology and immunohistochemistry, allowing to non-invasively establish the endotype of nasal polyps and to highlight all cytotypes, including mast cells, that cannot be visualized by the other two techniques. The recent demonstration of the close correlation between mast cell intraepithelial infiltrate and CRSwNP severity paves the way for new therapeutic possibilities aimed at reducing not only eosinophilic infiltration but also mast cell infiltration.

The role of biologics in the pathogenesis and treatment of eosinophilic esophagitis

Eosinophilic esophagitis (EoE) is a chronic immune-mediated disorder characterized by inflammation, and intraepithelial infiltration of eosinophils in the esophageal epithelium. It is associated with unbearable symptoms, such as dysphagia, and bolus impaction which affect the nutritional status, and quality of life of the patients. It is diagnosed from a thorough medical history, and confirmed by endoscopy and esophageal biopsy. Traditionally, it has been treated with elimination diet, proton pump inhibitors, and swallowed topical corticosteroids. However, several of the patients with EoE are refractory to the treatment. Eosinophilic esophagitis like eosinophilic asthma is a T helper type 2 lymphocyte (Th2)-mediated allergic disease. During an allergic inflammation Th2 cells secrete cytokines, such as interleukin-5 (IL-5). IL-4, and IL-13 which play a key role in the pathogenesis of EoE. Th2 cytokines promote eosinophilopoiesis, differentiation, proliferation, maturation and activation of eosinophils. Activated eosinophils liberate several cytotoxic cationic proteins, and reactive oxygen species which cause epithelial injury, inflammation, and remodeling, which later progresses to esophageal fibrosis. Dupilumab (dupixent®)is a monoclonal antibody which target the IL-4 receptor (IL-4Rα) which signals for both IL-4 and IL-13. Dupilumab is the only biologic which is effective, and approved for the treatment of EoE in adults and children 12 years and older. Treatment with dupixent® has been shown to improve dysphagia, dilate and relax the gullet, and allow patients to enjoy their cherished dish. Biologics such as dupilumab should be administered early in patients with EoE in order to prevent the menacing complications of the disease, such as esophageal stricture, and esophageal perforation, which necessitate surgical esophageal dilatation, and repair.

Eozinofilni ezofagitis

Eosinophilic esophagitis (EoE) is an inflammatory disorder characterized by intraepithelial eosinophilic infiltration followed by esophageal dysfunction. Pathophysiological mechanism of EoE is still not well understood, with several factors that may contribute, such as host immunity, environmental and genetic factors. Modern diagnostic for EoE should include: esophageal dysfunction, esophageal biopsy with at least 15 eosinophils per high power field and absence of other pathology characterized by increased number of eosinophils. The most common presenting symptoms in adults are dysphagia, heartburn, food impaction and chest pain. Children are usually presented with nausea and vomiting, anorexia, heartburn, regurgitation, chest burn and abdominal pain. Endoscopically, patients with EoE are characterized by longitudinal furrows (vertical lines, darker than surrounding mucosa), esophageal trachealization (numerous transversal rings, as in trachea), exudate (white plaques), edema (decreased mucosal vascularization), strictures and crepe-paper mucosa (mucosal friability and tearing during endoscopy). Histological features of EoE could be separated into major and minor criteria. Major criteria include: intraepithelial infiltration by eosinophils (&gt;15 eosinophils/HPF), eosinophilic microabscesses (≥4 eosinophils in a collection), eosinophils occupying outer layer of the squamous epithelium, epithelial sloughing and eosinophil degranulation. Minor criteria include: basal zone hyperplasia, lengthening of the epithelial papillae, intracellular edema and subepithelial fibrosis. In order to set adequate diagnosis, all other conditions related to increased number of eosinophils should be excluded. The most common and the most important differential is gastroesophageal reflux disease (GERD). Treatment of the EoE encompasses: proton pump inhibitors (PPI), corticosteroids (topical and systemic), elimination diet and esophageal dilation.

Клинико-морфологическая диагностика эозинофильного эзофагита

Eosinophilic esophagitis (EoE) is an immune-mediated disease that presents with dysphagia and esophageal bolus obstruction and is characterized by predominant intraepithelial eosinophilic infiltration at histology. The aim of our literature review was to delineate clinical and morphological EoE features in terms of differential diagnosis with other esophageal diseases, i.e., gastroesophageal reflux disease (GERD), inflammatory bowel diseases (IBDs), autoimmune diseases, and rare esophageal disorders. Clinical features of dysphagia and esophageal bolus obstruction and endoscopic criteria (according to EREFS) are typical of EoE. Nevertheless, eosinophilia of the esophageal mucosa is not a specific marker of mucosal injury and is not sufficient for EoE diagnosis. Therefore, eosinophilic esophagitis histological scoring system (EoEHSS) was developed. It involves assessment of intraepithelial eosinophilic infiltration, basal zone hyperplasia, eosinophilic abscesses, eosinophil surface layering, dilated intercellular spaces, surface epithelial alteration, dyskeratotic epithelial cells, and lamina propria fibrosis. Complex clinical, endoscopic, and histological evaluation allows accurate diagnosis of EoE and other causal factors of esophageal eosinophilia. Keywords: eosinophilic esophagitis, esophageal eosinophilia, gastroesophageal reflux disease, celiac disease, inflammatory bowel diseases, achalasia, atopy

MORPHOLOGICAL JUSTIFICATION OF IMMUNOREHABILITATION THERAPY OF CHRONIC NASOPHARYNGITIS ASSOCIATED WITH EBV

Adenotomy, the most widespread surgery in children, which does not solve the problem of symptoms in 19-26% patients. The main reasons are related diseases of nasopharynx, therefore, the indications for surgical intervention must be considered after evaluating the efficacy of conservative treatment. The relevant conservative treatment of CNP associated with EBV infection must include adequate immunorehabilitation effect on nasopharynx epitheliocytes, since the interaction of local immune response and cells infected with Epstein-Barr virus can lead to the control of virus infection.&#x0D; Objective of research: to establish the specific features of morphological changes in respiratory epithelium of pharyngeal tonsil in patients with chronic nasopharyngitis associated with EBV.&#x0D; Materials and methods. There was carried out the research of surgical material obtained as a result of endoscopic surgical intervention including the removal of hypertrophic part of pharyngeal tonsil in 15 patients diagnosed with chronic nasopharyngitis associated with EBV. The diagnosis was made on the ground of a typical clinical picture and determination of immunoglobulin M, G classes to Epstein-Barr virus in the blood.&#x0D; Results. It has been shown that intraepithelial lymphocytic and histiocytic infiltration can be determined morphologically and the edema of loose connective tissue, full blood vessels of the microcirculatory channel can be determined subepithelially which is the morphological sign of the chronic inflammation process.&#x0D; The identified structural reconstruction of epithelium reflects the development of virus-induced degenerative processes, namely, koilocytosis and spongiosis. The virus-induced changes in epithelium in the form of koilocytosis can be observed both in covering epithelia and in metaplastic epithelium, which demonstrates the persistence of viral infection in chronic inflammation.&#x0D; It has been found that on the background of chronic inflammation the cystic formations in the thickness of the lymphoid tissue are forming and the amphophilic masses of mucus can be visualized on the surface of epitheliocytes with hypersecretory changes, as well as similar mucous masses can be identified in the lumen of cysts of lymphoid tissue. The changes in epithelium in the form of koilocytosis can be seen in the epithelial crypts and cysts as well as indicating that the persistence of infection is viral.&#x0D; Conclusions:&#x0D; &#x0D; In CNP associated with EBV, the morphological changes of respiratory epithelium of nasopharynx are indications of both non-specific inflammatory processes (hyperplasia, metaplasia of epithelium, hypersecretory changes in cells, intraepithelial inflammatory cell infiltration), and specific for viral infections processes (koilocytosis, spongiosis, lymphocytes with wide cytoplasm).&#x0D; Structural changes are indications of the development of degenerative virus-induced processes (koilocytosis, spongiosis) combined with reactive changes in epithelial cells (hyperplasia, hypersecretion) that happen both in the surface epithelium and in the crypts with cyst formations.&#x0D; There can be seen phenomena of significant hypersecretory activities of epithelium including the accumulation of mucus on the surface of cells as well as in the lumen of crypts and cysts.&#x0D;

Respiratory epithelial adenomatoid hamartomas of the sinonasal tract: A histopathological analysis of 50 patients.

Respiratory epithelial adenomatoid hamartoma (REAH) is a benign lesion of the nasal cavity and paranasal sinuses. Here, we report the clinicopathological characteristics of REAH identified in 2065 cases with nasal/paranasal polypoid lesions treated with endoscopic sinus surgery (ESS) at our hospital from 2008 to 2021. Cases including the olfactory area were reviewed and 50 patients of REAH were identified pathologically (50/2065, 2.4%). The average age was 58.9 years old and the male/female ratio was 45/5. Grossly, REAH showed a whitish surface and elastic firm consistency. The histopathological characteristics included proliferation of small to medium-sized glands composed of ciliated respiratory epithelium containing goblet cells; thickening of the basement membrane compared to that for inverted papilloma (9.6 ± 2.4 vs. 1.3 ± 1.6 µm, p < 0.001); and no intra-epithelial neutrophilic infiltration. Among the REAH cases, 81% were associated with sinonasal inflammatory polyps. Many olfactory cleft polyps were REAH (38/98, 39%). The rate of REAH found in ESS in the last 7 years was higher than that in the first 7 years (3.17% vs. 1.62%, p = 0.032). Our results in Japanese patients are similar to those found in other countries, including male predominance. REAH is relatively common and that 39% of polyps taken from olfactory clefts are REAH.

Abstract P1-08-26: Morphologic characterization of tumor-infiltrating lymphocytes and its relation with pathological response in a series of breast cancer patients treated with primary chemotherapy

Abstract Introduction: Tumor-Infiltrating Lymphocytes (TILS) is a well-known predictor of response to primary chemotherapy1,2 and a prognostic factor for improved survival in different breast cancer subtypes3. We present data on the association of the level of TILs and morphologic characteristics of such infiltrate with pCR. Material and methods: A series of 477 breast cancer patients (479 tumors) treated with primary chemotherapy at Catalan Institut of Oncolgy - H.U. Bellvitge between January 2009 and December 2016. Chemotherapy consisted on anthracyclines and taxanes (plus trastuzumab if Her-2 positive disease). Levels of percentage of TILs using hematoxylin-eosin-stained sections of diagnostic core-needle biopsy were evaluated according to international guidelines in a prospectively defined retrospective analysis. Characterization of TILS consisted on identification of plasma cells, intraepithelial infiltrate vs stromal infiltrate and homogeneous vs heterogeneous infiltrate. A sub-classification using levels of TILS and heterogeneity was done for statistical purposes. Levels of TILS and their morphological characteristics were examined for their associations with pCR adjusted for predictive clinic-pathological factors, by univariate and multivariate logistic regression, statistical significance set at 0.05. A ROC curve was performed to look for a cut-point of TILs to predict PCR. Results: The mean value of TILs was 23.79% (SD, 24%). TILs were significantly higher in ductal carcinomas (39.1% vs 0%, p=0.007), grade 3 (55.2 vs 17.7%p&amp;lt;0.001), ki 67 &amp;gt;30 (48.8% vs 24.5%, p&amp;lt;0.001), HER2 (56.6%) and triple negative tumors (TNBC)(56.6%) vs luminal (20%), p&amp;lt;0.001). The level of TILS was set in 20% to predict pCR by a ROC curve (S=62.3% E=71%). Characterization of TILs found plasma cells in 41.1% of the samples, intraepithelial infiltrate in 11.2% and homogeneous infiltrate in 60.1%. Homogeneously high infiltrate was found in 20% of the samples. In the univariate analysis pCR was higher in samples with TILs &amp;gt; 20% (15.2% vs 41.5%, OR: 3.96 [95%CI, 2.57-6.10]; P &amp;lt; 0.001); plasma cells (OR 6.61 [95%CI, 1.51-28.8]; P=0.01), intraepithelial TILS (OR: 10.34 [95%CI, 2.22-48.01]; P =0.003), homogeneous high infiltrate (OR: 13.6 [95%CI, 3.04-60.77]; P =0.001). In luminal tumors, TILs over 20% predicted pCR (OR 12.3 [95%CI, 4.0-37.7]; P &amp;lt; 0.001) as well as in TNBC (OR 4.32 [95%CI, 1.77-10.53]; P=0 .001) but not in those cases with HER2 positive tumors (luminalB HER2 + HER2) (OR 1.65 [95%CI, 0.88-3.07]; P=0.118). In the multivariate analyses, levels of TILs &amp;gt; 20% were associated with higher pCR rates (adjusted odds ratio, 2.44 [95%CI, 1.48-4.01]; P &amp;lt; .001). Conclusions: The presence of TILs over 20% at diagnosis is an independent, positive, predictive marker of pCR in early breast cancer treated with neoadjuvant chemotherapy. Interestingly, the predictive information added by TILs &amp;gt;20% was higher in luminal and triple negative tumors compared to HER2 positive cases. Careful morphological characterization of TILS may add valuable predictive information and can be done in current pathologic laboratories with a well-trained breast cancer pathologist.

Characterization of immune cell infiltrate in tumor stroma and epithelial compartments in oral squamous cell carcinomas of Sudanese patients.

BackgroundTumor immune infiltrate has been explored in oral squamous cell carcinoma (OSCC), but studies on simultaneous characterization of multiple immune cell subtypes separately in stromal and intraepithelial tumor compartments are limited.ObjectivesWe aimed to investigate the immune cell infiltrate in OSCC by using immunohistochemistry (IHC) for a panel of inflammatory cells in stromal and epithelial tumor compartments for a better characterization of the tumors.MethodsThirty‐six OSCC lesions and nine normal oral mucosa (NOM) samples from patients attending Khartoum Dental Teaching Hospital, Sudan were investigated for presence of tumor infiltrating lymphocytes, tumor‐associated macrophages, tumor‐associated neutrophils, and PD‐L1 positive cells in the inflammatory infiltrate by single and double IHC. Digital quantitative analysis (Aperio Technologies Inc.) was performed separately for stromal and epithelial compartments.ResultsOSCC cases displayed a higher inflammatory infiltrate in the associated stroma, but not in the epithelial compartment when compared to NOM. The immunosuppressive type of inflammatory infiltrate, that is, T regulatory cells (FoxP3+ cells) was identified to be significantly higher in the epithelial compartment of tumors with advanced clinical state. An immunoscore developed by combining intraepithelial FoxP3+ and CD4+ cells was found significantly higher in lesions from elderly patients, localized at toombak dipping‐related sites, poorly differentiated OSCCs, or with loco‐regional lymph node spreading.ConclusionsDespite heavy immune cell infiltration in tumor‐associated stroma, the majority of OSCCs in this cohort displayed a low intraepithelial immune infiltration. An immunoscore based on combined CD4 and FoxP3 intraepithelial expression may serve as an indicator of advanced tumor progression and should be further investigated for its use as potential prognostic biomarker in OSCC.

Primary vulvar squamous cell carcinomas with high T cell infiltration and active immune signaling are potential candidates for neoadjuvant PD-1/PD-L1 immunotherapy

BackgroundA profound insight into the immune landscape of vulvar squamous cell carcinoma (VSCC) is lacking. Here, an in-depth interrogation of T cell infiltration, local immune contexture, signaling pathways and checkpoint...

The significance of focally enhanced gastritis in haematopoietic stem cell transplant recipients.

The histological diagnosis of acute gastric graft-versus-host-disease (aGVHD) in patients with a history of haematopoietic stem cell transplant (HSCT) is based on the presence of epithelial cell apoptosis and karyorrhectic debris. There is, however, limited information on the histological findings in patients who develop symptoms several months after transplant. Focally enhanced gastritis (FEG), defined by the presence of focal periglandular lymphohistiocytic inflammation with neutrophilic or lymphocytic intra-epithelial infiltration of gastric glands, has been described in patients with inflammatory bowel disease and in HSCT patients. The pattern closely resembles the focal periductal inflammation and lymphocytic exocytosis seen in chronic GVHD of the salivary gland. We sought to evaluate the significance of FEG in HSCT patients. Gastric biopsies from 151 HSCT patients who underwent endoscopies for GVHD-like symptoms were identified. Time from transplant to biopsy, presence of extra-gastric GVHD, medications and outcome were noted. Thirty-five biopsies showed FEG and 21 showed aGVHD; the remainder were either normal or showed non-specific changes. Twenty-one (60%) FEG patients had concurrent histologically proven extra-gastric GVHD. The time to biopsy in FEG patients was significantly longer than in aGVHD patients (162 versus 57days, P<0.01). Prior or subsequent gastric biopsies of 14 patients in the FEG cohort were also evaluated and, of these, six showed aGVHD while one showed persistent FEG. These findings suggest that FEG probably represents a form of late-occurring GVHD. This histological pattern should not be overlooked when identified in gastric biopsies from HSCT patients.

The 488th case: chronic diarrhea and abnormal liver function

A 56-year-old female was admitted to Department of Gastroenterology at Peking Union Medical College Hospital with diarrhea for seven months, and abnormal liver function for six months. She had a history of type 1 diabetes. The main clinical manifestations were recurrent fatty diarrhea and abnormal liver function, accompanied by abdominal and retroperitoneal lymphadenopathy, elevated CA19-9 and CEA. Progressive impairment of hepatic synthetic function and shrinkage of liver developed in a short period of time. The pathology of liver biopsy suggested that nodular regeneration of hepatocytes was followed by hyperplasia of thin bile ducts after submassive necrosis. Intestinal mucosa biopsies were performed twice. The pathology showed that the intestinal villi were completely blunt, accompanied with crypt hyperplasia. Goblet cells disappeared with reduced mucin. Paneth cells were barely seen without intraepithelial infiltration of lymphocytes. Rifaximin was not effective, while glucocorticoids improved clinical situation. The diagnosis of autoimmune enteropathy was finally confirmed by multidisciplinary team including departments of gastroenterology, pathology, endocrinology, hematology, infectious diseases, and rheumatology. With the administration of glucocorticoid and sirolimus, diarrhea relieved and liver function returned to normal.

Pembrolizumab in unresectable or metastatic MSI-high colorectal cancer: safety and efficacy

ABSTRACT Introduction Microsatellite instability-high (MSI-H) colorectal cancer (CRC) represents a unique subset of CRC characterized by elevated neoantigen expression and a high degree of intraepithelial T-cell infiltrate. These characteristics make MSI-H tumors particularly susceptible to immune checkpoint inhibitors (ICIs) such as pembrolizumab which inhibit the negative regulation of cytotoxic T-cells and promote T-cell mediated anti-tumor activity. Areas covered We discuss the drug development of pembrolizumab including the seminal studies which enabled the drug to garner FDA approvals in the refractory and first-line settings for patients with MSI-H CRC, the pharmacokinetic & pharmacodynamic profile of the agent, and the adverse event profile of the ICI. We also discuss unmet needs in the arena of ICIs including strategies to overcome tumor resistance and to increase the applicability of the agents to a broader population of CRC patients. Expert opinion Despite the anti-tumor activity of pembrolizumab in patients with MSI-H CRC, 30–35% of patients fail to derive any benefit. Ongoing research efforts are seeking to identify ICI combinations, which can overcome CRC resistance to pembrolizumab, move ICIs into the treatment paradigm for patients with localized MSI-H CRC and enable ICIs to become meaningful treatment options for patients with microsatellite stable CRC.

NK cell infiltration is associated with improved overall survival in solid cancers: A systematic review and meta-analysis

The immune landscape of a tumor is highly connected to patient prognosis and response to treatment, but little is known about how natural killer (NK) cells predict overall survival (OS) among patients with solid tumors. We present the first meta-analysis on NK cell infiltration into solid tumors as a prognostic indicator for OS, considering cancer types independently, and together. Samples were collected from 1973 to 2016 with results published between 1989 and 2020. From 53 studies, we found that NK cell infiltration corresponds with decreased risk of death (HR=0.34, 95% CI: 0.26–0.46; p<0.0001). Among studies that investigated the prognostic potential of NK cells in specific regions of the tumor, intraepithelial infiltration was better predictive of OS than NK infiltration in the tumor-adjacent stroma. Generally, NK cell infiltration is lower in advanced-stage and lower-grade tumors; nevertheless, it remains prognostically beneficial. This meta-analysis highlights an important prognostic role of NK cells in solid tumors, but exposes that few studies have considered the contributions of NK cells. Toward NK cell-based immunotherapies, it will be important to understand the conditions under which NK cells can be effective agents of tumor control.