NKT and NKT-like Cells in Autoimmune Neuroinflammatory Diseases-Multiple Sclerosis, Myasthenia Gravis and Guillain-Barre Syndrome.

NKT cells comprise three subsets-type I (invariant, iNKT), type II, and NKT-like cells, of which iNKT cells are the most studied subset. They are capable of rapid cytokine production after the initial stimulus, thus they may be important for polarisation of Th cells. Due to this, they may be an important cell subset in autoimmune diseases. In the current review, we are summarising results of NKT-oriented studies in major neurological autoimmune diseases-multiple sclerosis, myasthenia gravis, and Guillain-Barre syndrome and their corresponding animal models.

Lower weight gain after vaping cessation than after smoking quitting

Background: Smoking is frequently a way to control appetite and weight. The data concerning the body mass gain after quitting among the users of electronic cigarettes who have no prior history of smoking traditional cigarettes is inconsistent. Objective: In our study we have compared smoking and vaping impact on weight gain and glycaemia. Material and methods: 3 groups of rats were used. The group A was exposed to vapour and group B were exposed to smoke. Rats in the group C constituted the control group without nicotine exposition. Results: During 6 weeks of experiment weight gain of rats in the A and B groups was comparable, while animals from group C had gained signifi0cantly more. During 2 weeks after cessation of exposition to nicotine animals from group B gained more weight than rats of A and C group. Blood glucose was higher in group B than in groups A and C 24 h after last exposure to nicotine and 2 weeks after nicotine exposure cessation. Conclusion: Effects of vaping on weight increase is similar to smoking, but after vaping cassation weight gain is lower and comparable with nicotine nonusers.

CB2R agonist prevents nicotine induced lung fibrosis.

INTRODUCTION: Nicotine stimulates fibroblast proliferation while increasing inflammation and fibrosis of tissues. The cannabinoid receptor 1 (CB1R) is mainly located in the CNS, while cannabinoid receptor 2 (CB2R) is located in the immune cells within the body. CB2R regulates inflammatory processes and fibroblast function. PURPOSE: We investigated the impact of CB2R agonist, JWH 133 and the antagonist, AM630 on lung tissue, applied directly before nicotine application. MATERIAL AND METHODS: 40 mice were placed into 4 groups. The experimental groups received nicotine intraperitoneally at a dose of 0.05 mg/kg of body weight (BW) for 14 days. Group B also received AM630 (0.5mg/kg of BW), while Group A was administered with JWH133 (1 mg/kg of BW). Group N received nicotine alone. The Control group C received 0.9% NaCl. After decapitation, lung tissues were stained with H&E, Trichrome Masson's method, and IHC against CTGF and α-SMA. The digital image processing system Image J with the IHC profiler plugins was then employed, optical density and IHC optical density score were calculated. RESULTS: In the N group, an increase in the thickness of alveolar spaces (9.16 SD4.95µm vs. 4.77SD2.99µm in the C group), leukocytes infiltration and collagen deposition has been observed(OD: 0.20 SD0.0vs 0.07SD0.04 in the C group). In the B group, the alveolar space thickness has been the highest (11.57SD8.13µm). Furthermore, in this group, hyperaemia, destruction of lung structure, hyperplasia of II type pneumocyte and interstitial fibrosis has been observed (OD: 0.23 SD0.08). In contrast, the lung tissue of the A group has had normal structure and the thinnest alveolar septum (3.88 SD2.64µm). The expression of CTGF and α-SMA has been the highest in the B group. CONCLUSION: Nicotine induces interstitial lung fibrosis that is enhanced by the CB2R antagonist and diminished by the CB2R agonist. Therefore, the CB2R agonist may offer a protection against fibrosis.

Decrease in Lipid Droplets in Adrenal Cortex of Male Wistar Rats after Chronic Exposure to Energy Drinks.

Background and objectives: Energy drinks are popular non-alcoholic beverages. They are consumed in large amounts, mainly by active, young people. Although they are easily accessible and marketed as safe, numerous cases of adverse effects have been published, including cardiac arrest, arrythmias, acute hepatitis, and renal failure. The aim of the current study is the assessment of energy drink influence on the histological structure of adrenal cortex in rats. Material and Methods: 15 male young Wistar rats were equally divided into three groups: control (C), experimental (E) and reversibility control (RC). C group received water and standard rodent food ad libitum while both E and RC groups had additionally unlimited access to energy drinks. C and E groups were decapitated after 8 weeks and RC was given another 8 weeks without energy drinks. Adrenal glands were embedded in paraffin blocks and 5 µm slides were prepared and stained according to standard H&E and Masson's trichrome protocols. Additionally, immunohistochemical stainings against Ki-67, p53, CTGF and caspase-3 were prepared. Results: Decreased vacuolization and numerous pyknotic nuclei were noted in E and RC groups. Overexpression of caspase-3 was noted both subcapsular in zona glomerulosa and along sinusoids in zona fasciculata. Increased collagen deposition in zona glomerulosa and zona fasciculata of E and RC was observed. Insular and irregular overexpression of CTGF was noted. The overall picture of CTGF expression matched the Masson's trichrome. No significant difference was observed in Ki-67 expression. Conclusions: The results of the current study suggest that the stimulation is so intense that it causes significant damage to adrenal cortical cells, resulting in their apoptosis. It seems, however, that the observed effects are at least partially reversible.

Tobacco smokers and electronic cigarettes users among Polish universities students.

BACKGROUND: Electronic cigarettes (e-cigarettes) are small battery-powered electronic devices, heating the liquid to produce vapour--in most cases the latter contains nicotine and several flavourings. E-cigarettes are highly advertised across the media, mainly as healthy substitute to conventional cigarettes, aid in quitting smoking addiction or way of circumventing ban on smoking in public places. OBJECTIVE: The aim of study was obtaining epidemiological data on cigarette smoking and electronic cigarette usage among Polish universities students. MATERIAL AND METHODS: Students of different Polish state universities were asked to fill a self-prepared survey on cigarette-smoking and electronic cigarette usage. 1068 fulfilled questionnaires were gathered. The population was divided into two subgroups--medical universities' students (n=545) and non-medical universities students (n=523). RESULTS: 23.78% of respondents declared current smoking while 57.0% admitted ever smoking. The mean duration of smoking among current smokers was 4.17±2.53 years. 56.30% of current smokers tried quitting at least once. 31.46% of students declared ever using e-cigarettes (37.28% (n=195) among non-medical universities' students and 25.87% (n=141) among medical universities' students and 8.33% current usage. Among the latter 52.81% admitted simultaneous smoking. 26.97% of current e-cigarettes' users declared having experienced side effects of e-cigarettes. 42.70% (n=456) of respondents viewed e-cigarettes as safer than conventional cigarettes, this group comprises of 40.54% (n=212) non-medical and 44.77% (n=244) medical universities' students. 85.39% (n=912) of students viewed e-cigarettes as generally unhealthy, there were 83.56% (n=437) non-medical and 87.16% (n=475) medical universities' students among this group. CONCLUSIONS: The frequency of e-cigarettes usage resembles current status in many Western countries. Collected data shows high frequency of e-cigarettes usage and conventional cigarettes smoking among students (also medical universities' students). The situation requires intensive preventive measures to limit and reduce the popularity of tobacco products along with modern equivalents like electronic cigarettes.

Expression of CD226 on γδ T cells is lower in advanced chronic lymphocytic leukemia and correlates with IgA, IgG and LDH levels.

BACKGROUND: Gamma-delta (γδ) T cells comprise an important subset of human T cells, responding to viral and bacterial infections, and are significant for cancer immunosurveillance. Human γδ T cells are divided into 5 major subsets, namely Vδ1-Vδ5, of which the latter 3 have limited available literature. At present, Vδ2 is the most studied subpopulation. OBJECTIVES: In the current paper, we focused on non-Vδ2 cells in chronic lymphocytic leukemia (CLL). We assessed the expression of co-inhibitory checkpoint receptors (CTLA-4, PD-1 and TIGIT) and co-stimulatory (CD226 and NKp30) molecules separately on Vδ1 and Vδ3-Vδ5 cells. MATERIAL AND METHODS: We assessed γδ T cells for their expression of both cytotoxicity-related (NKp30, CD226) and co-inhibitory (PD-1, TIGIT) molecules with flow cytometry in CLL patients. Moreover, we evaluated the expression of TIGIT and CD226 ligand (PVR , CD155) in neoplastic B cells in CLL patients with quantitative real-time polymerase chain reaction (qPCR). RESULTS: A significant accumulation of Vδ1 T cells was noted, while no difference was observed in the total percentage of Vδ2 cells. Contrary to our initial hypothesis, the impact of CLL burden on CD226 and TIGIT expression was lower than anticipated. The former tends to be lower in more advanced disease. Finally, a strong upregulation of CD155 (PVR) was noted on CLL-derived B cells when compared to healthy B cells. CONCLUSIONS: Chronic lymphocytic leukemia regulates the expression of the CD155-CD226/TIGIT axis. Contrary to expectations, the ligand is significantly more affected than the receptors. Nevertheless, the relatively high expression of CD155 and TIGIT makes CLL an interesting target for anti-TIGIT immunotherapy.

The Mysterious Actor-γδ T Lymphocytes in Chronic Lymphocytic Leukaemia (CLL).

Chronic lymphocytic leukaemia (CLL) is the most common leukaemia among adults. It is the clonal expansion of B cells expressing CD19 and CD5. Despite significant progress in treatment, CLL is still incurable. γδ T cells comprise an important subset of the cytotoxic T cells. Although γδ T cells in CLL are dysfunctional, they still can possibly be used for immunotherapy. The current paper reviews our understanding of γδ T lymphocytes in CLL.

PECAM-1 Is Down-Regulated in γδT Cells during Remission, but Up-Regulated in Relapse of Multiple Sclerosis.

Introduction. PECAM-1 and NKRP1A are both involved in the vascular transmigration of T lymphocytes. Vascular transmigration is a crucial process in multiple sclerosis pathogenesis. Methods and aim. The current paper presents an analysis of PECAM-1 and NKRP1A expression on γδ T cells. Expression of PECAM-1 and NKRP1A on subsets of γδ T cells was performed with flow cytometry. Results. Based on the flow cytometry data, PECAM1 was slightly differentially modulated on γδ T cells-it was up-regulated during relapse, but down-regulated during remission. Moreover, a significant downregulation of CD3 expression was noted on γδ T cells from MS patients, most notably during relapse. Conclusions. This may be a sign of the overall activation of γδ T cells in the course of multiple sclerosis.

Changes in Histological Structure, Interleukin 12, Smooth Muscle Actin and Nitric Oxide Synthase 1. and 3. Expression in the Liver of Running and Non-Running Wistar Rats Supplemented with Bee Pollen or Whey Protein.

Introduction: Bee pollen is a natural substance obtained from flowers by bees. It is a rich source of protein, vitamins and minerals. It can be used as a dietary supplement. Bee pollen has been investigated for the treatment of some diseases with promising potential. It can be helpful in supportive therapy for dyslipidemia, metabolic syndrome, diabetes type 2, as well the prevention and control of coronary heart disease and myocardial infarction. Whey protein is a rich source of amino acids. It is a basic dietary supplement for many athletes, both professional and amateur. It stimulates muscle growth and provides nutrition for cachectic patients. Aim of the study: The objective of the study was to assess the impact of dietary supplementation of bee pollen or whey protein on the Wistar rat liver histological structure and expression of interleukin 12, smooth muscle actin and nitric oxide synthases among running and non-running rats. Material and methods: Thirty male Wistar rats were divided into six equal groups, three running and three non-running. Among both there was one control, one supplemented with bee pollen and one receiving whey proteins. After 8 weeks, all animals were decapitated and their livers were collected. Five micrometer thick slides were prepared and used for classical histological staining and immuno-histochemistry. ImageJ image analysis software was used to measure optical density and immunohistochemistry profile coverage. Results: Among all groups, morphology of liver was similar. In the running control group, expression of interleukin-12 (IL-12) was decreased as well as expression of endothelial nitric oxide synthase (eNOS) in a group of bee pollen supplemented rats. No significant changes in α- smooth muscle actin (α-SMA) expression was observed. Conclusions: Bee pollen is proving to be a questionable choice for athletes as an alternative to whey protein. Bee pollen supplementation affects hepatocyte cellular activity and has hepatoprotective effects. Whey protein performs worse in this regard. Lower antioxidant properties were found in groups supplemented with bee pollen than with whey protein.

γδ T Lymphocytes in Asthma: a Complicated Picture.

A minor subset (approximately 5%) of peripheral T cells has their TCR build up from γ and δ chains instead of α and ß-those are the γδ T lymphocytes. They can be functionally divided into subsets, e.g., Th1-, Th2-, Th9-, Th17-, Tfh-, and Treg-like γδ T cells. They share some specifics of both innate and adaptive immunity, and are capable of rapid response to a range of stimuli, including some viral and bacterial infections. Atopic diseases, including asthma, are one of major health-related problems of modern western societies. Asthma is one of the most common airway diseases, affecting people of all ages and having potential life-threatening consequences. In this paper, we review the current knowledge about the involvement of γδ T cells in the pathogenesis of asthma and its exacerbations. We summarize both the studies performed on human subjects as well as on the murine model of asthma. γδ T cells seem to be involved in the pathogenesis of asthma, different subsets probably perform opposite functions, e.g., symptom-exacerbating Vγ1 and symptom-suppressing Vγ4 in mice model of asthma.

IL-15 Is Overexpressed in γδ T Cells and Correlates with Disease Severity in Relapsing-Remitting Multiple Sclerosis.

Interleukin 15 (IL-15) is known to be involved in the pathogenesis of multiple sclerosis (MS). An animal study revealed a distinct subset of IL-15-producing γδ T cells that correlate with disease severity. The aim of the current study was to test whether such a subset is also present in humans and its importance for the pathogenesis of MS. The peripheral blood from 29 patients with relapsing-remitting MS (including 6 relapses) and 22 controls was stained with monoclonal antibodies and analyzed with flow cytometry. The existence of IL-15+ γδ T cells was confirmed. Moreover, the percentage of IL-15+ γδ T is significantly increased in MS patients and correlates with disease severity. Nevertheless, additional functional studies are needed to fully understand the importance of those cells in multiple sclerosis pathogenesis.

γδ T lymphocytes in the pathogenesis of multiple sclerosis and experimental autoimmune encephalomyelitis.

The aim of the current review is to summarize the results of studies on the role of γδ T cells in the pathogenesis of multiple sclerosis and its animal model - the experimental autoimmune encephalomyelitis. Despite the fact that numerous studies have been performed, the role of γδ T is still not fully understood. It seems that there are two distinct subpopulations - one exacerbating the disease (IL-17-producing) and the other playing a protective role (IFN-γ-secreting). Nevertheless, future studies are required for an understanding of γδ T cells role in multiple sclerosis.

Changes in histological structure and nitric oxide synthase expression in aorta of rats supplemented with bee pollen or whey protein.

Various protein-based supplements are at least periodically consumed by 30%-40% of sportspeople. The current study compares cardiovascular effects of diet supplementation with 2 different protein-rich products: bee pollen and whey protein. Thirty Wistar rats were divided into 2 groups, one subjected to daily moderate physical activity and one not. Each group consisted of 3 subgroups: control, whey-protein-supplemented, and bee-pollen-supplemented. After 8 weeks, rats were decapitated, and proximal parts of thoracic aortas were collected and embedded in paraffin blocks. Histological slides were stained according to standard hematoxylin and eosin, Masson's trichrome, and Verhoeff - Van Gieson staining. Special immunohistochemical stains against neuronal nitric oxide synthase (nNOS), endothelial nitric oxide synthase (eNOS), and alpha smooth muscle actin were also prepared. Histological evaluation revealed noticeable changes in all supplemented groups: disturbances in elastic laminae, slight increase in collagen deposition, and significantly lowered nNOS and eNOS expression. The prevalence of small atherosclerotic plaques was the highest in non-running supplemented groups, while in running supplemented groups it resembled the prevalence in control groups. Both running groups had thinner tunica media than control. Both supplements exert visible effects on aortic structure, but the difference between them is far less evident. In some aspects, however, the bee pollen seems to be even slightly more harmful, which may be related to various possible contaminants like mycotoxins or pesticides.

RORγT is overexpressed in iNKT and γδ T cells during relapse in relapsing-remitting multiple sclerosis.

The aim of the current study is to evaluate IL-17 production and RORγT, and IL-23R expression by iNKT, Th17 and γδ T cells in the peripheral blood of relapsing-remitting multiple sclerosis patients. Samples of peripheral blood from 21 relapse patients and 12 remission patients, and 15 healthy volunteers were stained with monoclonal antibodies for flow cytometry analysis. No significant differences in iNKT, γδ T and Th17 percentages were noted. The significant overexpression of RORγT was observed in all three subpopulations - therefore, iNKT, γδ T and Th cells may be an important source of IL-17 shortly prior to the relapse.