Lymphoid Component Research Articles

Immune Profiles of Myeloma Tumor Microenvironment May Predict Sensitivity or Resistance to Elotuzumab

Background: Elotuzumab (ELO), an immunobiologic therapy targeting SLAMF7/CD319, is effective in the treatment of multiple myeloma (MM) with clinical indications in relapsed/refractory disease in combination with lenalidomide or pomalidomide. ELO binds SLAMF7 on the surface of MM cells to increase immune recognition while also binding SLAMF7 on NK cells resulting in the activation of an effector cell population capable of readily killing via antibody-dependent cellular cytotoxicity (ADCC) mechanisms. ELO efficacy has also been linked to enhanced antibody-dependent cellular phagocytosis (ADCP). Given the dependence of ELO activity on immune elements of the tumor microenvironment (TME), we hypothesized that characterization of the immunologic constituents of the immune TME (iTME) would provide cues capable of predicting clinical efficacy.Methods: Nineteen patients were enrolled on a single-center clinical trial run at Moffitt Cancer Center comparing clinical activity of ELO in combination with low-dose (10mg) vs high-dose (25mg) lenalidomide and dexamethasone in patients with biochemical relapse while on lenalidomide maintenance after first line therapy. Bone marrow aspirate (BMA) and peripheral blood (PB) samples we collected at baseline, after completion of 2 treatment cycles, and at time of progression. Cells isolated from the BMA were analyzed by multiparameter flow cytometry (MPFC) using 4 panels; 3 panels characterized lymphocytes according to maturation, activation and polarization (T H1, T H2, T H17 or T Reg) and a 4 th panel characterized myeloid elements. Data was acquired on a BD Symphony cytometer and analysis was performed using FlowJo software.Results: Patient samples were categorized according to initial therapeutic response. Responders (n=6) achieved >PR at best response, Progressors (n=5) were identified as patients whose disease markers increased >25% and Stable Disease (n=8) was identified as demonstrating <25% change in serologic disease markers. Here, we present a comparison of phenotypic profiles from the BM samples acquired at baseline before treatment initiation from patients classified as Responders or Progressors (patients demonstrating stable disease as best response were analyzed separately). Sample analysis by MPFC revealed distinct populations uniquely associated with responses as well as with treatment refractoriness. Principal component analysis (PCA) of the myeloid immune compartment shows 2 clusters representing distinct populations detectable only in Responders, while another large cluster, also expressing a granulocytic phenotype consistent with granulocytic myeloid suppressor cells, is prevalent in the Progressor patients but nearly absent from Responder patients, as might be predicted. Similarly, PCA identified monocyte/macrophage clusters uniquely associated with Responder or Progressor samples. Evaluation of lymphoid populations using 3 lymphoid panels also reveals distinctive clustering patters by PCA highlighting populations differentially associated with Responder or Progressor samples. Notably, while NK cells are present in all patient samples, there is a population of immature NK cells enriched in Responder patients that is largely absent from the Progressor population.Conclusion: Immune profiling of the myeloid and lymphoid components of the baseline BM iTME in MM prior to treatment with ELO reveals distinct patterns of cellular constitution associated with responsiveness or refractoriness to subsequent treatment. While NK cells are known to be important to the ELO mechanism of action, our data suggests that ELO activity is compromised in the absence of immature NK cells. This work suggests that measurable characteristics of the cellular components of the iTME may provide important cues to help predict ELO therapeutic efficacy. These observations will need to be confirmed in a larger data set. DisclosuresShain: Amgen Inc: Consultancy, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Sanofi Genzyme: Consultancy, Speakers Bureau; Adaptive Biotechnologies Corporation: Consultancy, Speakers Bureau; AbbVie: Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis Pharmaceuticals Corporation: Consultancy; BMS: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; GlaxoSmithLine, LLC: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen oncology: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Karyopharm Therapeutics Inc.: Honoraria, Research Funding. Baz: BMS, sanofi, Karyopharm, Janssen, AbbVie: Consultancy, Research Funding; Oncopeptides: Consultancy; GlaxoSmithKline: Consultancy, Honoraria; Merck: Research Funding. Nishihori: Novartis: Research Funding; Karyopharm: Research Funding.

Primary Warthin’s-like adenocarcinoma of the lung: A clinicopathological, immunohistochemical, and molecular analysis of three cases

Three cases of primary pulmonary adenocarcinoma with prominent lymphoid stroma and papillary features mimicking Warthin’s tumor are presented. The patients are two women and one man ages 52, 62, and 74 years respectively. Clinically, the patients presented with non-specific symptoms of cough, dyspnea, and chest pain. Imaging showed the presence of an intrapulmonary mass in the right lower, right upper and left lower lobe. All patients underwent lobectomy. Histologically, the tumors were characterized by the presence of and oncocytic papillary growth pattern embedded in a lymphoid rich background. Immunohistochemical stains for TTF-1, keratin 7, and beta-catenin were positive in the epithelial component, while CD20 showed strong positive staining in the lymphoid component. In addition, CD4 and CD8 also showed positive staining in a ratio of 3–4:1, EBER was negative. Kras mutations with wild type EGFR were identified in one case. Clinical follow-up ranging from 8 to 24 months was obtained showing that all patients are alive without recurrence. The cases herein presented represent an unusual histological variant of primary lung adenocarcinoma, which closely mimics Warthin’s tumor.

Treatment-Free Remission: the New Goal in CML Therapy.

Purpose of ReviewTreatment-free remission (TFR) is considered one of the main goals of therapy in patients with CML. Our goal in this paper is to review the current data on TFR, and discuss future directions.Recent FindingsMultiple studies have demonstrated that attempting a treatment-free remission is safe and effective in a select group of patients. More recent data suggested that undetectable BCR-ABL1 by digital PCR prior to discontinuation is highly predictive of successful TFR. However, some patients have a successful TFR with no evidence of clinical disease despite persistent detectable BCR-ABL1. Some recent studies have shed some more light on possible mechanisms for this phenomena. Some possible mechanisms include immune mechanism, BCR-ABL1 detected in the lymphoid component only, or stem cell exhaustion.SummaryTFR should be discussed with patients with CML. Patients who achieve a sustained deep molecular response may be eligible to attempt TFR, however, setting expectations that overall only 20% of patients with newly diagnosed CML will achieve a successful TFR. The importance of compliance to treatment early on cannot be overemphasized. Further studies using other drugs to get patients to a deeper remission in order to be eligible for TFR attempt, or attempting a second TFR in patients who had disease recurrence after first TFR attempt, are currently underway.

Delayed maturation of thymic epithelium in mice with specific deletion of \u03b2-catenin gene in FoxN1 positive cells

Wnt signalling pathways have been reported to be involved in thymus development but their precise role in the development of both thymic epithelium (TE) and thymocytes is controversial. Herein, we examined embryonic, postnatal and adult thymi of mice with a specific deletion of β-catenin gene in FoxN1+ thymic epithelial cells (TECs). Together with a high postnatal mouse mortality, the analysis showed severe thymic hypocellularity, largely due an important reduction in numbers of developing thymocytes, and delayed, partially blocked maturation of mutant TECs. Affected TECs included largely cortical (c) TEC subsets, such as immature MTS20+ TECs, Ly51+ cTECs and a remarkable, rare Ly51+MTS20+MHCIIhi cell subpopulation previously reported to contain thymic epithelial progenitor cells (TEPCs) (Ulyanchenko et al., Cell Rep 14:2819–2832, 2016). In addition, altered postnatal organization of mutant thymic medulla failed to organize a unique, central epithelial area. This delayed maturation of TE cell components correlated with low transcript production of some molecules reported to be masters for TEC maturation, such as EphB2, EphB3 and RANK. Changes in the thymic lymphoid component became particularly evident after birth, when molecules expressed by TECs and involved in early T-cell maturation, such as CCL25, CXCL12 and Dll4, exhibited minimal values. This represented a partial blockade of the progression of DN to DP cells and reduced proportions of this last thymocyte subset. At 1 month, in correlation with a significant increase in transcript production, the DP cell percentage increased in correlation with a significant fall in the number of mature TCRαβhi thymocytes and peripheral T lymphocytes.

Collision of Epstein\u2013Barr virus-positive and -negative gastric cancer, diagnosed by molecular analysis: a case report

BackgroundEpstein–Barr virus (EBV)-positive gastric carcinoma (GC) is defined by the proliferation of GC cells with EBV infection. The co-existence of EBV-positive and -negative components in a single GC is rare. We report a case of GC with the co-existence of EBV-positive and EBV-negative components, in which we performed—for the first time—various molecular analyses to elucidate their histogenesis.Case presentationAn 81-year-old man was diagnosed with GC based on the results of endoscopy and a pathological examination of the biopsy specimen. Systemic chemotherapy was performed, since lymph node and lung metastases were diagnosed based on computed tomography. Total gastrectomy and lymph node dissection were performed after chemotherapy, after confirming that the size of the metastatic lymph nodes had decreased and that the lung metastasis had disappeared. Grossly, a type 3 tumor was located in the middle posterior part of the stomach body. At the cut section, the tumor consisted of a white and solid part on the anal side of the tumor and a flat and elevated part on the oral side. Histologically, the former part consisted of GC with lymphoid stroma and the latter part was composed of poorly differentiated adenocarcinoma without prominent lymphocytic infiltration. The two histopathological components were clearly separated from each other. On EBV-encoded small RNA (EBER)-in situ hybridization (ISH), the part with the lymphoid stroma component was positive, while the other part was negative. Immunohistochemistry revealed that both components showed the overexpression of p53. Sequencing of TP53 using DNA extracted from the two components was conducted, and revealed different patterns. Targeted next generation sequencing revealed MYC amplification in the EBV-positive component of the tumor and HER2 amplification in the EBV-negative part. Immunohistochemistry revealed that the EBV-positive part was C-MYC( +)/HER2(−) and the EBV-negative part was C-MYC(−)/HER2( +). Correspondingly, chromogenic ISH and dual-color ISH showed amplification of C-MYC and no amplification of HER2 in the EBV-positive part, and no amplification of C-MYC and amplification of HER2 in the EBV-negative part.ConclusionWe presented a case of collision of two different GCs composed of EBER-ISH ( +)/C-MYC ( +) and EBER-ISH (−)/HER2 ( +) cells.

The potential role of follicular helper T cells and helper T cells type 1 in Warthin tumour

Warthin tumour (WT) is a benign tumour of the salivary gland that proliferates in both glandular epithelial and lymphoid tissue components, and rarely exhibits cystic changes. T follicular helper cells (Tfh) are involved in the formation and maintenance of germinal centres, the differentiation of B cells into plasma cells, and the maintenance of helper T cell type 2 (Th2)-dominant humoral immune responses. T-bet induces differentiation into helper T cell type 1 (Th1) by suppressing differentiation into Tfh and enhances cellular immune responses. The objective of this study was to enhance our understanding of the immune responses and relationship between Tfh and Th1 cells in patients with WTs. In this study, we classified WTs (n = 64) into solid-type (n = 25) and cyst-type (n = 39). We also performed immunostaining of the Tfh markers CXCR5 and CD40 L, and the Th1 marker T-bet for statistical analysis. The cyst-type exhibited significant atrophy of the germinal centre area (P = 0.0019), significantly fewer Tfh-positive lymphocytes in germinal centres (P < 0.0001), and significantly more T-bet-positive lymphocytes in the epithelium (P = 0.0017). We observed that Tfh were involved in the formation and maintenance of lymphoid follicles in WTs. In the cyst-type, Th2-dominant humoral immune responses were suppressed, and Th1-dominant cellular immune responses may have caused damage to tumour tissue.

Immune System in Action.

Tumor exists as a complex network of structures with an ability to evolve and evade the host immune surveillance mechanism. The immune milieu which includes macrophages, dendritic cells, natural killer cells, neutrophils, mast cells, B cells, and T cells is found in the core, the invasive margin, or the adjacent stromal or lymphoid component of the tumor. The immune infiltrate is heterogeneous and varies within a patient and between patients of the same tumor histology. The location, density, functionality, and the crosstalk between the immune cells in the tumor microenvironment influence the nature of immune response, prognosis, and treatment outcomes in cancer patients. Therefore, an understanding of the characteristics of the immune cells and their role in tumor immune surveillance is of paramount importance to identify immune targets and to develop novel immune therapeutics in the war against cancer. In this chapter, we provide an overview of the individual components of the human immune system and the translational relevance of predictive biomarkers.

Micronodular Thymomas With Prominent Cystic Changes: A Clinicopathological and Immunohistochemical Study of 25 Cases.

Twenty-five cases of micronodular thymomas with prominent cystic changes are presented. The patients are 13 men and 12 women between the ages of 38 and 69 years. Clinically, the majority of patients presented with nonspecific symptoms of cough, chest pain, and dyspnea. Four patients were asymptomatic. Diagnostic imaging showed the presence of an anterior mediastinal mass and surgical resection of the tumor mass was performed in all the patients. Histologically, all the tumors were characterized by the presence of cystic structures of varying sizes lined by different types of epithelium. In addition, the tumors were characterized by nodules of epithelial cells embedded in a lymphocyte-rich stroma with germinal centers. Twenty-one tumors were encapsulated while 4 tumors were minimally invasive. Immunohistochemical stains were positive for keratin in the nodular epithelial component while CD45 and CD20 were positive in the lymphoid component. Clinical follow-up ranging from 12 to 24 months was obtained in 19 patients. All the patients were alive and well without disease. No clinical follow-up was available in 6 patients. The cases presented in this article highlight the existence of cystic micronodular thymomas, which can be easily misdiagnosed as a multilocular thymic cyst.

Inflammatory lobular hemangioma: A vascular proliferation with a prominent lymphoid component. Review of a series of 19 cases

In the last 30 years, there has been a strong interest in vascular proliferations. Pyogenic granuloma was not only renamed lobular capillary hemangioma, but also the conceptual interpretation was also changed from an overgrowth of granulation tissue to a genuine hemangioma (or benign vascular neoplasm). We describe 19 cases of patients who presented clinically with a vascular lesion, characteristically a pyogenic granuloma or lobular hemangioma, where the histopathological findings led to the pathologic concern for a lymphoma of the skin. These benign lesions with a dense lymphoid infiltrate were further defined on the basis of different vascular and lymphoid immunohistochemical markers as inflammatory lobular hemangiomas. We propose that given the considerable histopathological overlap between acral pseudolymphomatous angiokeratoma, T-cell rich angiomatoid polypoid pseudolymphoma of the skin, and other designations of some of these vascular proliferations with a rich and dense lymphoid infiltrate, they might constitute a spectrum of vascular lesions with varying clinical presentations.

Low Molecular Weight Cytokeratin Immunohistochemistry Reveals That Most Salivary Gland Warthin Tumors and Lymphadenomas Arise in Intraparotid Lymph Nodes

The nature of the lymphoid stromal component in Warthin tumor and lymphadenoma of the parotid gland has been a controversial topic in salivary gland pathology for decades. Two theories exist: first, that these tumors arise from salivary gland inclusions within intraparotid lymph nodes; and second, that they arise within salivary gland parenchyma and induce tumor-associated lymphoid proliferation (TALP). A recent study demonstrated that low molecular weight cytokeratin is effective in distinguishing salivary gland tumors within lymph nodes from those inducing TALP via identification of extrafollicular reticulum cells, which are only found in true lymph nodes. Twenty-one Warthin tumors and 4 lymphadenomas were retrieved from the archives of the Department of Pathology at University of Texas Southwestern Medical Center.Cam5.2 immunohistochemistry was performed on each case and independently evaluated by two pathologists. Extrafollicular reticulum cells were identified by Cam5.2 immunostaining in 21 of 21 Warthin tumors (100%), and 3 of 4 lymphadenomas (75%). Extrafollicular reticulum cells were consistently localized to the perisinusoidal and paracortical areas of the lesions studied. Extrafollicular reticulum cells were identified via low molecular weight cytokeratin Cam5.2 immunohistochemistry in all of the Warthin tumors and most of the lymphadenomas evaluated.This finding strongly supports the notion that these most if not all of these tumors arise within intraparotid lymph nodes, presumably from salivary gland inclusions entrapped during embryonic development.

Myeloid and lymphoid sinchronous neoplasm with t(8;13) (p12;q12) - FGFR1 in a positive SARS-CoV-2 patient: Case report

33 year-old male with a myelo-lymphoid neoplasm presentation at the debut. He went to Hospital because of fever and throat pain, with SARS-CoV2 infection at the same time. Initial blood tests showed hyperleukocytosis (49,320 x 10^3µL) and high cell count of neutrophils (39,440x10^3µL). Peripheral blood studies were suggestive of myeloproliferative Syndrome, and SARS-CoV-2 test was positive. Flow citometry (FCM) studies showed lymphocytes with patological inmunophenotype (CD3-) indicative of lymphoid component. Genetic studies showed 8p11/FGFR1 traslocation with caryotipe: 46,XY,t(8;13)(p11.2;q12)[22]/46,XY[3]. Myeloid genetical studies were negative, but TCR was positive. We concluded he presented lymphoid / myeloid complex neoplasm with significant tumor burden at debut, which is characteristic of this traslocation. Bone marrow and peripheral blood studies were compatible with myeloproliferative Neoplasm, even though genetic studies guide to lymphoid disease. Both ganglio biopsy and FCM of bone marrow diagnosed a T-lymphoma. B symptoms (asthenia and weight loss) and palpable lymphadenopaties also indicate T component. This complex hematological neoplasm was aggravate by SARS-CoV-2 infection. It was decided to treat the haematological disease once the infection was solved. Patient received standard doses according to usual protocol for T acute lymphoblastic leukemia (T-ALL) in Spain, in an alogenic stem cell trasplant (ASCT) candidate. Also SARS-CoV2 infection was treated and solved. Now patient is responding to treatment, showing antibodies to the virus. With control-test negatives until today doing every two weeks, recommended by Infectious disease department of our center. grow factor receptor (FGFR) is a non-exclusive molecule of human being. Four different types of FGFR are known in our environment. FGFR tirosin kinase activity has a N-terminal sequence codified by FIM gene, which joins the kinase domain. This covalent bond is responsible for oncogenic potential of this hematopoietic cells. The gen receptor plays an important role in Hematopoiesis. Its hyperactivity leads to abnormalities in signal pathways, resulting with cellular mutations or Hematological neoplasms.1 Abnormalities in this factor are also responsible for genetic diseases such as Pffeifer or Kallman syndrome (FGFR3). Also tumors (FGFR1- lymphoblastic lymphomas) or genetic traslocations related to Hematological malignancies are described. They are usually associated to chromosome eight traslocation (t8;13), as a result of gene fusion ZMYM2(ZNF198)-FGFR1. Few cases are registered about 8p11/FGFR1 traslocation-related neoplasms at literature. Mixed myelo-lymphoid neoplasms are usually related to this traslocation.2 This neoplasms associated with FGFR1 mutation frequently get the origin in Stem cell, resulting in a wide and heterogeneus clinical presentation.3

Abstract 5522: Evaluation of eribulin-induced alterations of the intact immune cell landscape in spleens and tumors from tumor-bearing immunocompetent mice

Abstract Eribulin, a synthetic analog of the marine sponge natural product halichondrin B, is approved in the US for certain patients with advanced breast cancer or liposarcoma. Its mechanisms of action include both cytotoxic, antimitotic effects as well as non-cytotoxic effects on tumor vasculature, phenotype and microenvironment. Known aspects of eribulin's effects on tumor phenotype, including reversal of epithelial-mesenchymal transition (EMT) and induction of cellular differentiation in various preclinical models, led us to speculate that eribulin might also alter immune homeostasis in the in vivo setting, both in tumors themselves as well as supporting immune organs such as the spleen. Here, we report on eribulin's effects on both lymphoid and myeloid lineage cellular components in both spleen and tumors from immunocompetent mice bearing subcutaneous Lewis lung carcinoma (LLC) isografts. Surprisingly, spleens of vehicle treated, LLC tumor-bearing mice are approximately twice as large spleens from non-tumor-bearing mice, with an excellent correlation (r2 = 0.92) between individual tumor size and spleen weight arguing for a direct cause and effect relationship between the presence of tumor and splenomegaly. Tumor-induced splenomegaly is associated with increased proliferation of splenic non-lymphoid (CD4-/CD8-/CD19-) cells, including both CD11b+ mature myeloid lineage cells as well as CD11b- cells. Due to high levels of P-glycoprotein (PgP) characteristic of most murine tumors, LLC tumors respond poorly to eribulin, a known PgP substrate. Thus, 3 doses of MTD eribulin (1.6 mg/kg; Q4Dx3) led to only a 30% reduction in tumor volume relative to vehicle-treated mice. Unexpectedly, however, despite this reduced tumor volume, spleens from eribulin-treated mice were twice again larger than the already-enlarged spleens from vehicle-treated tumor-bearing mice, and the clear relationship between tumor size and spleen weight was abolished. This additional eribulin-induced splenomegaly was associated with enhanced proliferation of the same CD4-/CD8-/CD19- non-lymphoid cells, CD11b+ myeloid lineage cells and CD11b- cells as in vehicle-treated, tumor-bearing mice. Evaluation of immune cell components in the associated fixed tumors from eribulin- and vehicle-treated mice is currently ongoing to assess what effect, if any, such changes in the spleen will have on the tumor immune microenvironment; these results will be presented at the meeting. In conclusion, our results point to significant and unexpected effects of eribulin treatment on the immune cell landscape in spleens and tumors from tumor-bearing immunocompetent mice. Citation Format: Haochen Wu, Keyi Zhu, Xiaolong Tu, Diana Albu, Mary Woodall-Jappe, Bruce A. Littlefield. Evaluation of eribulin-induced alterations of the intact immune cell landscape in spleens and tumors from tumor-bearing immunocompetent mice [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5522.

Cellular reactions of the mucous membrane of the intestinal system after uranium incorporation

A pilot study, which was performed on white laboratory male rats and included a morphological study of the mucous membrane of the jejunum and colon, revealed the long-term effect of a single oral exposure to an aqueous solution of depleted uranium oxides; this supported dynamic changes in the studied criteria determining metabolic, regenerative, and local regulatory processes in one, three and six months. A lymphoid component appeared to be the criterion of high radio sensitivity and a marker of nonspecific protection indicating deterioration of homeostasis. In the time dynamics of long-term periods, a change in all the parameters was detected when using immunohistochemical, histochemical, and specific methods at the level of the epithelium lining the relief formations of the mucous membranes of the intestinal system organs. The oxidative stress resulting in three months after exposure to depleted uranium, which was evidenced by a significant decrease in the content of anti-apoptotic Bcl-2 protein in the epithelial cells of the jejunely mucosa. Signs of its delayed effect after six months in the colon induced the pronounced nature of the reactions of stromal cell components that determined patterns of protective mechanisms of the intestinal-associated immune system with a greater expressiveness in the jejunum. The factual material cited in the work suggests the existence of a homeostatic immune response in the mucous membrane of the small and large intestines that controls cell proliferation, as a special form of immune surveillance of the state of cytodifferentiation.

Combined Effects Of Unloading And Radiation On Skeletal Muscle In Mice

In preparation for upcoming space missions to the Moon and Mars, there is a need to understand how space stressors (e.g. microgravity, radiation) affect different physiological systems. As skeletal muscle is a critical organ, not only for locomotion but also for overall body homeostasis, defining the molecular impact of microgravity and radiation on this tissue will help developing new, or fine-tuning current countermeasures to maintain health and physiological function of space travellers. PURPOSE: To investigate the effects of combined radiation and unloading on anabolic/catabolic and immune/inflammatory processes on skeletal muscle in mice. METHODS: Ten C57/BL6J mice were subjected to 14-d hind-limb unloading by tail suspension with an acute radiation session (dose=25 mGy, X-ray) on day 7 of unloading (HLUR). Ten mice were used as control (CTRL; similar cages, sham radiation). Mice were sacrified and soleus muscle was immediately dissected, weighed and frozen. Then, RNA was extracted and converted to cDNA. Gene expression of anabolic/catabolic (i.e. myostatin, MuRF-1, Atrogin-1, PGC-1α) and immune/inflammatory markers (i.e. CD4, CD8, IFNγ, CD11b, MHCII, TNF, IL-6) was assessed by RT-PCR. Independent t-tests were use to compare HLUR vs. CTRL. RESULTS: Soleus muscle weight was ~30% lower in HLUR vs. CTRL (P<0.001). Myostatin expression was greater in HLUR vs. CTRL (1.8-fold, P=0.014). MHCII expression was higher in HLUR vs. CTRL (2.4-fold, P<0.001). There was a trend for group differences (P<0.08) in CD11b and TNF mRNA content with HLUR showing greater values than CTRL. Gene expression of CD4, CD8 and IFNγ was barely detected in either group. CONCLUSIONS: The combination of unloading and radiation has a major impact on skeletal muscle. Apart from inducing muscle atrophy, as indicated by the decreased muscle weight and increased myostatin levels, these two space stressors altered the immune profile within the muscle. The increased gene expression of MHCII and CD11b indicates that the myeloid component of the immune system is activated upon unloading and radiation in skeletal muscle. In contrast, the almost undetected mRNA levels of CD4, CD8 and IFNγ may imply that unloading and acute radiation have little impact on the lymphoid component. These findings should be followed up with immunohistochemical analysis.

Cytopathology and diagnostics of Warthin's tumour.

Warthin's tumour (WT) is a benign epithelial salivary tumour, one type of salivary adenoma. Histologically, WT is structured of two components, epithelial tissue that often lines cystic formations and lymphoid tissue in the tumour stroma. FNA is a reliable diagnostic approach in the diagnosis of salivary gland lesions allowing a highly accurate categorization of benign tumour-like lesions, benign tumours and malignant tumours. In the proposed Milan reporting system of salivary gland lesions, WT is categorized in the IVA group of benign neoplasms. Accurate cytological diagnosis is straightforward when three characteristic components are present: oncocytes, either isolated or associated in clusters, lymphocytes and lymphoid cells and often an inflammatory/necrotic-like substance. Also, specific features of scintigraphy and radiological imaging contribute to the diagnosis of WT. WT is categorized according to Seifert G. et al in 4 types, depending on the proportions of the epithelial component and lymphoid stroma. Differential cytopathological and pathohistological diagnosis include other salivary gland lesions with lymphoid, oncocytic epithelial and cystic components. In some cases, such as the metaplastic WT variant, there are additional cytopathological and histological diagnostic difficulties. Moreover, bilateral, multicentric or multiple and infrequently seen extra-salivary localizations of WT are associated with further cytopathological diagnostic difficulties. Also, a rare possibility of malignant transformation of the epithelial or lymphoid component of WT as well as possible association with other primary tumours remains a challenge in accurate cytopathological and histological diagnosis of WT.

Parathyroid adenoma with prominent lymphocytic infiltration: A potential pitfall in frozen sections and cytology reporting

Introduction: Parathyroid adenoma with prominent lymphocytic infiltration is rare, with only 12 cases reported in the English scientific literature. All these cases were not related to synchronous autoimmune or IgG4-related disease and presented clinically with primary hyperparathyroidism. Case presentation: A 65-year-old woman presented with hypercalcemia and a neck mass, suggestive of parathyroid adenoma. She underwent parathyroidectomy and histopathological examination of the specimen showed a hypercellular parathyroid nodule comprising mature lymphoid cells admixed with parathyroidal chief cells. A comprehensive immunohistochemistry panel confirmed the benign nature of the lesion. The patient remains asymptomatic a year after the surgery. Discussion: Parathyroid adenoma with prominent lymphocytic may be misdiagnosed as a lymph node at frozen section or cytology smears by an unsuspecting pathologist or cytologist, if the lymphoid component is predominant on the slide examined. Additionally, the morphology could be misinterpreted as a metastasis of parathyroid carcinoma into a peri-thyroidal lymph node. For this distinction, correlation with clinical and radiological findings is crucial and clues to adenoma include presence of a rim of uninvolved parathyroid parenchyma, low mitotic activity (<5%) and positive staining for parafibromin, bcl-2 and MDM2 on immunohistochemistry.

Composite Epstein-Barr virus-positive mucosa-associated lymphoid tissue lymphoma and Epstein-Barr virus-negative diffuse large B-cell lymphoma in the parotid salivary gland of a patient with Sj\xf6gren\u2019s syndrome and rheumatoid arthritis: a case report

BackgroundEpstein-Barr virus is associated with many human hematopoietic neoplasms; however, Epstein-Barr virus-positive mucosa-associated lymphoid tissue lymphoma is extremely rare. In routine clinical practice, detection of mucosa-associated lymphoid tissue lymphoma and diffuse large B-cell lymphoma in a tissue sample presumes a clonal relation between these neoplasms and that diffuse large B-cell lymphoma developed by transformation of the mucosa-associated lymphoid tissue lymphoma. However, evidence to support this presumption is sparse and controversial. Assessment of the clonal relationship of the lymphoid components of a composite lymphoma is important for understanding its pathogenesis and correct diagnosis.Case presentationWe present an unusual case of composite lymphoma (Epstein-Barr virus-positive mucosa-associated lymphoid tissue lymphoma/Epstein-Barr virus-negative diffuse large B-cell lymphoma) in the parotid salivary gland of a 62-year-old Caucasian woman with Sjögren’s syndrome and rheumatoid arthritis. Simultaneous occurrence of mucosa-associated lymphoid tissue lymphoma and diffuse large B-cell lymphoma in the parotid salivary gland led us to initially assume a clonal relationship between diffuse large B-cell lymphoma and mucosa-associated lymphoid tissue lymphoma. Epstein-Barr virus was detected by in situ hybridization and polymerase chain reaction in the mucosa-associated lymphoid tissue lymphoma, but not in diffuse large B-cell lymphoma, suggesting that these lymphomas were not clonally related. Fragment analysis of frame region 3 polymerase chain reaction products from microdissected mucosa-associated lymphoid tissue lymphoma and diffuse large B-cell lymphoma components revealed different clonal pattern rearrangements of the immunoglobulin heavy chain gene.ConclusionsOur patient’s case highlights the importance of assessing the clonal relationships of the lymphoid components of a composite lymphoma and Epstein-Barr virus screening in mucosa-associated lymphoid tissue lymphoma in patients with autoimmune disease.

Overview of Basic Immunology and Clinical Application.

Tumor exists as a complex network of structures with an ability to evolve and evade the host immune surveillance mechanism. The immune milieu which includes macrophages, dendritic cells, natural killer cells, neutrophils, mast cells, B cells, and T cells are found in the core, the invasive margin, or the adjacent stromal or lymphoid component of the tumor. The immune infiltrate is heterogeneous and varies within a patient and between patients of the same tumor histology. The location, density, functionality, and cross-talk between the immune cells in the tumor microenvironment influence the nature of immune response, prognosis, and treatment outcomes in cancer patients. Therefore, an understanding of the characteristics of the immune cells and their role in tumor immune surveillance is of paramount importance to identify immune targets and to develop novel immune therapeutics in the war against cancer. In this chapter, we provide an overview of the individual components of the human immune system and the translational relevance of predictive biomarkers.

Vector vaccine Flu/ESAT-6 application in the regimen of complex therapy of drug-sensitive and drug-resistant experimental tuberculosis estimation of the efficiency

The steady growth of drug-resistant tuberculosis, charac-terized by low therapeutic efficacy, requires the develop-ment of innovative approaches to the treatment of this infection. One of such approaches is vaccine therapy that has immunostimulating activity and is capable to correct dysfunction of the immune system in this infection. In this study, the authors aimed to estimate the efficacy of the mucosal vector vaccine Flu/ESAT-6 included into the complex therapy of experimental pulmonary tuberculosis. C57BL/6 mice were intravenously infected with drug-sus-ceptible (DS) (M. tuberculosis H37Rv) or drug- resistant (DR) (M. tuberculosis Beijing genotype) MBT strains. DS-in-fection was treated with two anti-TB drugs (HR), and DR-infection with four (AETppBq). Flu/ESAT-6 vaccine based on the attenuated influenza A virus strain expres-sing the mycobacterial protein ESAT-6 was administered intranasally at a dose of 6 lg/mouse. Efficacy of vari-ous therapeutic regimens was assessed with biometric, bacteriological, and histological methods. Two optimal therapeutic regimens of Flu/ESAT-6 administration were chosen: twice (with a 3 week interval) at the beginning of chemotherapy in case of DS infection; and thrice (with a 4 week interval) 2 weeks after the beginning of chemo-therapy in case of DR infection. Flu/ESAT-6 addition to the anti-tuberculosis treatment led to a significant increase in the efficacy of therapy for both forms of infection which manifested in (i) the pulmonary bacterial load decrease at the end of a 2-months therapy of DS tuberculosis by 1.83 lg CFU; and after 4-months therapy of DR tuberculosis by 0.93 lg CFU; (ii) decrease in the prevalence of specific and the severity of exudative inflammation, and reduction of alternative components in case of DR tuberculosis; (iii) stimulation of the lymphoid component of the cellular im-mune response in lungs, demonstrated by reorganization of granulomas’ cellular composition to lymphoid type, and in the formation of peribronchial lymphohistiocytic infiltrates. Thus, therapeutic immunization with Flu/ESAT-6 vaccine significantly increases the efficacy of experimen-tal tuberculosis therapy and can shorten the duration of anti-tuberculosis therapy which is especially important in case of DR tuberculosis characterized by a slow involution of specific inflammation.

Risk of malignancy in “atypia of undetermined significance” category of salivary gland fine‐needle aspiration: A bi‐institutional experience

Many prior institutional and multi-institutional studies have applied the Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) retrospectively to their specimens to determine the risk of malignancy (ROM) of each category. Most of these studies focused on general assessment of the system and risk classification. However, there seems to be less focus on the category of atypia of undetermined significance (AUS) that could be attributed to the low number of cases that could fit into this category. Herein, we present a bi-institutional experience with this category. A computerized search of the databases was performed to identify all salivary gland fine-needle aspiration (FNA) in two institutions over a period of 12 years. The final diagnosis of each case was reclassified based on MSRSGC, and histology follow-up was retrieved. Sixty AUS cases (out of 1560 salivary gland FNA) were identified with a rate of 3.8%. Forty cases (66%) had a subsequent tissue material. Correlation with histology revealed that the estimated ROM is 37.5% (15/40) and the overall ROM is 25% (15/60). Fifty percent of the cases had a prominent lymphoid component and most commonly represented lymphomas, reactive lymph node or sialadenitis. The AUS category is a heterogeneous group of lesions with predominant lymphoid-rich entities. Some variability exists between institutions with most having higher ROM than the suggested 20% by the MSRSGC atlas.