Relationship between lymph concentration of cytokines and structural transformations in mesenteric lymph nodes in chemotherapy, surgical treatment, and chemotherapy of experimental breast cancer

Цель исследования - анализ корреляции морфометрии брыжеечных лимфатических узлов и концентрации цитокинов в лимфе грудного протока при химиотерапии рака молочной железы, хирургическом лечении и последующей химиотерапии. Методика. Рак молочной железы индуцировали введением N-метил-N-нитрозомочевины 5 раз с интервалом 7 сут подкожно в область 2-й молочной железы справа. Курс химиотерапии проходил по схеме CMF. Корреляцию между концентрациями 24 цитокинов лимфы и числом клеток структурных зон лимфатических узлов оценивали по коэффициенту ранговой корреляции Спирмена. Результаты. После химиотерапии РМЖ, по сравнению с РМЖ без лечения, морфологические преобразования в лимфатических узлах свидетельствуют о снижении активности местного иммунного ответа. Исследование корреляции концентрации цитокинов в лимфе со структурными изменениями в лимфатических узлах выявило зависимости направленные на повышение иммуномодулирующего и противоопухолевого действия цитокинов. После оперативного лечения РМЖ и последующей химиотерапии, по сравнению только с химиотерапией РМЖ, выявлены положительные связи иммунобластов с цитокином GRO/KC в герминативных центрах, цитокина IL-6 - с митотически делящимися клетками в герминативных центрах и мозговых тяжах, IL-5 - с иммунобластами в мозговых тяжах, хемокина MIP-1a - со зрелыми плазматическими клетками в мозговых синусах. Увеличено количество иммунобластов, средних и малых лимфоцитов в герминативных центрах, возросло количество малых лимфоцитов, незрелых и зрелых плазматических клеток в мозговых синусах. Увеличены площади мозговых тяжей и паракортикальной зоны. Выявлена корреляция: цитокина IL-1α с малыми лимфоцитами, IL-6 с иммунобластами, IL-7 и IL-18 - со средними лимфоцитами, GRO/KC - с иммунобластами, IL-17 - с макрофагами в Т-зависимой зоне; IL-7 и IL-18 - с иммунобластами, IL-12 - с макрофагами, MIP-1a и MCP-1 со зрелыми плазматическими клетками в мозговых синусах. Заключение. После оперативного лечения РМЖ c последующей химиотерапией, по сравнению только с химиотерапией РМЖ, выявлены взаимозависимости концентрации цитокинов в лимфе грудного протока с морфологическими изменениями в брыжеечных лимфатических узлах, которые могут указывать на повышение активности местного звена иммунного ответа. The aim of this study was to analyze correlations of the morphometry of mesenteric lymph nodes with cytokine concentrations in thoracic duct lymph in chemotherapy and surgical treatment with subsequent chemotherapy of breast cancer. Methods. Breast cancer was induced by subcutaneous injection of N-methyl-N-nitrosourea 5 times with 7-day intervals, into the region of the 2nd breast on the right. The course of chemotherapy was performed according to the CMF scheme. Correlations between concentrations of 24 cytokines of the lymph and cells of lymph node structural regions were estimated by the Spearman rank correlation coefficient. Results. After chemotherapy for breast cancer compared to untreated breast cancer, morphological transformations in lymph nodes indicated decreased activity of the local immune response. Analysis of correlations between lymph concentrations of cytokines and structural changes in lymph nodes identified relationships aimed at increasing the immunomodulatory and antitumor effects of cytokines. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, positive correlations were found for immunoblasts with cytokine GRO/KC in germinative centers, for cytokine IL-6 with mitotically dividing cells in germinative centers and medullary cords, for IL-5 with immunoblasts in medullary cords, and for chemokine MIP-1a with mature plasma cells in medullary sinuses. Numbers of immunoblasts and medium and small lymphocytes were increased in germinative centers whereas numbers of small lymphocytes and immature and mature plasma cells were increased in medullary sinuses. Areas of medullary cords and the paracortical zone were increased. Correlations were found for cytokine IL-1α with small lymphocytes, for IL-6 with immunoblasts, for IL-7 and IL-18 with medium lymphocytes, for GRO/KC with immunoblasts, for IL-17 with macrophages in the T-dependent zone, for IL-7 and IL-18 with immunoblasts, for IL-12 with macrophages, and for MIP-1a and MCP-1 with mature plasma cells in medullary sinuses. Conclusion. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, cytokine concentrations in lymph of the thoracic duct were observed to correlate with morphological changes in mesenteric lymph nodes, which may indicate increased activity of the local immune response.

Spatiotemporal Expression of Foxl2 and Dmrt1 before, during, and after Sex Determination in the Sea Turtle Lepidochelys olivacea

The sex of sea turtles is determined by temperature during egg incubation. Thus, climate change affects the sex ratio, exacerbating their vulnerability to extinction. Understanding spatiotemporal effects of temperature on sex determination at the gonadal level may facilitate the design of strategies to mitigate the effects of global warming. Here, we used qRT-PCR and immunofluorescence to analyze the spatiotemporal expression of <i>Dmrt1 </i>and <i>Foxl2</i> in developing gonads of <i>Lepidochelys olivacea</i> incubated at male-producing temperature (MPT, 26°C) or female-producing temperature (FPT, 33°C). Although both transcription factors are expressed in bipotential gonads up to stage 25, the timing of their sexually dimorphic regulation differs. Whereas the dimorphic expression of Dmrt1 protein initiates at stage 24, Foxl2 protein was expressed specifically in females at stage 25. Interestingly, whereas Dmrt1 colocalizes with Sox9 in cell nuclei of primary medullary cords to form the testis cords, Foxl2 protein is first detected in Sox9-negative cells of primary medullary cords, prior to its substantial expression in the ovarian cortex. Thus, results suggest that the temperature-dependent regulation of sexual pathways is stochastic among the cells of primary medullary cords in undifferentiated bipotential gonads of the olive ridley.

Гистоархитектоника паренхимы лимфатических узлов млекопитающих с различными типами строения внутриузлового лимфатического русла

<p>The article analyzes the features of the histoarchitectonics of the lymph nodes of the bull (Bos taurus) and the pig (Sus scrofa domestica), depending on the type of structure and localization of the intranodal lymphatic channel. We studied somatic (Limphonodi (L.) cervicales superficiales) and visceral (L. jejunales) lymph nodes of clinically healthy mature male bulls and swine (16 and 6 months old, respectively). A complex of classical histological techniques was used, as well as the method of impregnating sections of lymph nodes with silver nitrate, modified by the authors. The main accumulative-distribution link in the lymph nodes of the bull is the subcapsular sinus (type I lymphatic collector), and in the lymph nodes of the pig - the capsular (intratrabecular) lymphatic tanks (type II lymphatic colector). In nodes with collectors of type I, the cortex has a simple layered structure, its outer layer is formed by a compact cortical plateau (interfollicular zone), and the inner layer is formed by a complex of spherical units of the deep cortex. In nodes with collectors of type II, the cortex is layered-folded, uneven in width. Cortical folds are formed along the capsular trabeculae with intratrabecular lymphatic tanks. The cortex plateau in the layered-folded cortex is more developed at the base of the folds, and the units of the deep cortex are at their apexes, where they form clusters in the form of specific nest-shaped structures. In nodes, regardless of the type of intranodal lymphatic channel, the surface cortex (cortical plateau) is located directly under the underlying lymphatic collectors, repeating their shape, the zones of clonal proliferation of B-lymphocytes are formed along the main collector on the basis of cortical plateau and its derivative structures (on the basis of paracortical and medullary cords). The zones of proliferation of T-lymphocytes are maximally close to the main collector, separated from it by a strip of cortical plateau, form a complex of spherical thickenings, which together form a deep cortex. The stroma and parenchyma are more developed in the nodes with collectors of type II (cumulative relative volume of stroma – 9-14% and 6-10%, parenchyma – 80-87% and 70-81%, respectively), and lymphatic sinuses - in nodes with collectors of I type (13-20% and 4-6% respectively). In the parenchyma of the lymph nodes of both groups, the zones of proliferation of T-lymphocytes predominate (the centers of deep cortex units are 27-42%), as well as the zones of accumulation of plasma cells and antibody formation (medullary cords – 17-29%), the first of which are more developed in the pig, and the latter at the nodes of the bull. The cumulative relative volume of the interfollicular zone (cortical plateau) in the studied nodes does not exceed 6-11%, and the zone of clonal proliferation of B lymphocytes (lymph nodulus) is 5-14%. These zones are more developed in the lymph nodes of the pig. Paracortical cords have the minimum and practically equivalent relative volume in the nodes of both groups (3-5%). The study shows that the principles of localization of the main specialized cellular zones in the lymph nodes of the domestic bull and the pig are universal, and the histoarchitectonics of the parenchyma has a clearly expressed specificity. Features of the histoarchitectonics of the parenchyma and its quantitative characteristics are determined by the type of structure of the intranodal lymphatic channel (the character of the localization and spatial configuration of the main accumulative-distributive unit of the nodes). </p>

The compartments of the parenchyma of the lymph nodes in newborn bull calves of domestic cattle (Bos taurus)

The article analyzes the features of the structure of the lymphoid lobules of the parenchyma of the superficial somatic (Limphonodi subiliaci, L. cervicales superficiales), profund somatic (L. axillares proprii L. poplitei), somatovisceral (L. iliaci mediales, L. retropharyngei mediales) and visceral (L. mediastinales caudales, L. ileocolici) lymph nodes of newborn bull calves of domestic cattle. To visualize clearly the boundaries of the structural components of lymphoid lobules we used the author’s modification of the impregnation of total median frozen histological sections with silver nitrate. We have established a high level of tissue differentiation of the lymph nodes, a significant development of the lymphoid parenchyma, the division of the parenchyma into lymphoid lobules, the presence in the lobules of all the main structural components that are represented by two morphotypes. The first morphotype is ribbon-like perisinusoidal cords (interfollicular zone, paracortical and medullary cords). The second morphotype is rounded lymphoid formations (central zones of deep cortex units, lymphatic nodules). Lymphoid lobules are located along the marginal sinus in one row, they are better developed and differentiated in the visceral lymph nodes. In all the lymph nodes, the lymphoid lobules have a similar histoarchitectonic, and each structural component of the lymphoid lobules has a specific architectonic of the reticular meshwork and the density of the location of the fibroblastic reticulocytes. We determined that the structures of the first morphotype which provide the migration of lymphocytes, the detection of antigens and the accumulation of plasmocytes are more developed. We have established that the relative volume of structures of the first morphotype is 4.5–8.0 times larger than the volume of the structures of the second morphotype, which provide clonal proliferation of T and B lymphocytes, especially in deep somatic lymph nodes. Among the zones of the second morphotype, predominate T-dependent zones, the relative volume of which considerably exceeds the volume of B-dependent zones (lymphoid nodules): in the superficial somatic lymph nodes by 14–30 times, profound somatic by 12–14 times, somatovisceral by 6–7 times and visceral by 4.5–5.5 times. We determined that lymphatic nodules can form in different parts of compartments: in the interfollicular zone and paracortical cords of all lymph nodes and in the medullary cords of the visceral lymph nodes. The study shows that the parenchyma of the lymph nodes of newborn bull calves has a high degree of maturity, contains a full set of structural markers of immunocompetence, among which predominate the components that support lymphocyte migration, antigen detection and accumulation of plasma cells.

Distribution of Porcine Haemal Nodes and Morphological Variations in the Haemal Nodes of Cattle and West African Dwarf Goats

Apparently healthy, non-lactating, non-gravid adult large white pigs (15), adult cattle (10) and adult West African Dwarf (WAD) goats of either genders were used to investigate the morphology of haemal nodes using gross anatomical and histological techniques. The results demonstrated that the haemal nodes of pigs occurred in the thoracic, abdominal and pelvic regions along the course of blood vessels. The ranges of the longest diameters of the dark brown organs were 6.1 – 30.3 cm in pigs, 4.1 – 18.7 cm in cattle and 2.7 – 8.2 cm in WAD goats. Bovine haemal nodes showed cortical and medullary parenchymal areas, with the cortex demonstrating primary and secondary lymphoid follicles in a circumferential fashion. The medulla showed blood filled medullary sinusoids and medullary cords. In WAD goat, the reticular cells and smooth muscle cells of the capsule were oriented in different directions and the subcapsular, cortical and medullary sinusoids exhibited erythrocytes in pockets. In conclusion, the present study has provided information on the distribution of haemal nodes in pigs as well as other unique morphological features of cattle and WAD goat haemal nodes that could aid their identification and suggest their roles in the body.

Distinctive localization of antigen-presenting cells in human lymph nodes

Professional antigen-presenting cells (APCs) are sentinel cells of the immune system that present antigen to T lymphocytes and mediate an appropriate immune response. It is therefore surprising that knowledge of the professional APCs in human lymph nodes is limited. Using 3-color immunohistochemistry, we have identified APCs in human lymph nodes, excluding plasmacytoid APCs, that fall into 2 nonoverlapping classes: (1) CD209+ APCs, coexpressing combinations of CD206, CD14, and CD68, that occupied the medullary cords, lined the capsule and trabeculae and were also scattered throughout the diffuse T-lymphocyte areas of the paracortex; and (2) APCs expressing combinations of CD1a, CD207, and CD208, that were always restricted to the paracortex. Surprisingly, this second class of APCs was almost entirely absent from many lymph nodes. Our data suggest that most CD208+ cells, often referred to as “interdigitating cells,” derive from migratory APCs, and that the major APC subset consistently resident in the paracortex of human lymph nodes is the CD209+ subset. All APC subsets were demonstrated to be in close contact with the fibroreticular network. The identification of 2 distinct APC populations in the paracortex of human lymph nodes has important implications for understanding T-lymphocyte responses and optimizing vaccine design.

Expression of mRNAs encoding oestrogen receptor (ER) α and ERβ, androgen receptor and progesterone receptor during gonadal and follicular development in the marsupial brushtail possum (Trichosurus vulpecula)

The objective of the present study was to determine which ovarian cells express mRNAs for oestrogen (ERα and ERβ), androgen (AR) and progesterone (PR) receptors during ovarian and follicular development in the brushtail possum. Expression of ERα and/or ERβ mRNA was observed from birth, initially in cells of the blastema, then in the medullary cords from Day 20. ERα was expressed in the oocytes and granulosa cells of secondary and antral follicles. Preovulatory follicles did not express ERα mRNA, although their oocytes were not examined for any gene. ERβ mRNA was observed in oocytes at all follicular stages examined, but was not consistently observed in granulosa or theca cells. Expression of AR mRNA before Day 40 was very faint; thereafter, expression was observed in the medullary cords, peaking between Days 60 and 120. Oocytes, granulosa cells and theca of secondary and antral, but not preovulatory, follicles expressed AR mRNA. PR mRNA was expressed throughout the gonad by Day 20. Granulosa cells of some secondary and antral follicles and theca of antral follicles expressed PR mRNA. Thus, the expression of mRNAs encoding steroidogenic receptors in a time- and cell-specific manner supports a role for steroids in the process of ovarian follicular formation and growth.

Dendritic Cell Populations in Colon and Mesenteric Lymph Nodes of Patients With Crohn's Disease

Dendritic cells (DCs) are key cells in innate and adaptive immune responses that determine the pathophysiology of Crohn's disease. Intestinal DCs migrate from the mucosa into mesenteric lymph nodes (MLNs). A number of different markers are described to define the DC populations. In this study we have identified the phenotype and localization of intestinal and MLN DCs in patients with Crohn's disease and non-IBD patients based on these markers. We used immunohistochemistry to demonstrate that all markers (S-100, CD83, DC-SIGN, BDCA1-4, and CD1a) showed a different staining pattern varying from localization in T-cell areas of lymph follicles around blood vessels or single cells in the lamina propria and in the MLN in the medullary cords and in the subcapsular sinuses around blood vessels and in the T-cell areas. In conclusion, all different DC markers give variable staining patterns so there is no marker for the DC.