Immunomodulatory Activity and Phytochemical Profile of Infusions from Cleavers Herb

Extracts from aerial parts of G. aparine (cleavers) constitute a herbal remedy with monography in British Herbal Pharmacopeia. On the European market, there are several drugs and food supplements consisting of Galium extracts. In folk medicine, cleavers was used topically in Europe, Asia, and the Americas to treat skin diseases. In several remedies, cleavers is also listed as an immunomodulatory active herb influencing the defense response of the human body. The aim of this study was to investigate the immunostimulatory activity and antioxidant potential in vitro of a raw infusion of cleavers and bioactive fractions. The functional activity of lymphocytes in the reaction of the lymphocyte blast transformation (RLBT) method was used for immunomodulatory activity assays and direct scavenging of 2,2-diphenyl-1-picrylhydrazyl (DPPH), nitric oxide (NO), and hydrogen peroxide (H2O2) was chosen for the examination of antioxidant activity. It was shown that both the raw extract and fractions show significant immunostimulatory and scavenging activities. The obtained data partially justify the traditional use of cleavers as topical remedy for skin infections and for wounds.

Phytochemical Profiles and In Vitro Immunomodulatory Activity of Ethanolic Extracts from Galium aparine L.

Galium aparine L., family Rubiaceae, is a widely spread species in the Galium genus. The herb of G. aparine is part of folk remedies and dietary supplements. In this study, we analyzed the chemical composition and immunomodulatory activities of G. aparine herb ethanolic extracts obtained from the plant material by maceration with 20%, 60% or 96% ethanol. The contents of hydroxycinnamic acid derivatives, flavonoids and polyphenols were determined spectrophotometrically, with extractives and polysaccharides quantified gravimetrically. The qualitative composition was studied using UHPLC-DAD-MS/MS analysis; isolation not previously described in G. aparine quercetin rhamnoglucoside was carried out through column chromatography, and the immunomodulatory activity of extracts was determined in the reaction of lymphocyte blast transformation. Major constitutes of extracts were iridoids, i.e., monotropein, 10-desacetylasperulosidic acid and asperulosidic acid; p-hydroxybenzoic acid; hydroxycinnamic acid derivatives, i.e., 3-O-caffeoylquinic, 5-O-caffeoylquinic, 3,4-O-dicaffeoylquinic, 3,5-O-dicaffeoylquinic, 4,5-O-dicaffeoylquinic acids and caffeic acid derivatives; flavonoids, i.e., rutin, quercetin 3-O-rhamnoglucoside-7-O-glucoside, and isorhamnetin 3-O-glucorhamnoside. Significantly, quercetin 3-O-rhamnoglucoside-7-O-glucoside was first isolated and identified in Galium species so far investigated. All G. aparine herb ethanolic extracts stimulate the transformational activity of immunocompetent blood cells, with 96% ethanolic extract being the most active. The data obtained necessitate further research into the mechanisms of immunomodulatory activity of extracts from G. aparine herb.

Cell-mediated immunity in visceral obesity

Visceral obesity is associated with the development of several diseases, such as osteoarthritis (OA), cardiovascular disease (CVD), type 2 diabetes mellitus (DM), and breast cancer (BC). Recent years have discussed the value of the immune system in the pathogenesis of the disease, but its role remains unclear. The aim of the study was to investigate the cellular immune responses in diseases comorbid with visceral obesity (OA, CVD, type 2 DM, BC). The study included 36 patients with OA, 61 – CVD, 60 - type 2 DM, 98 - breast cancer in postmenopausal women, 30 patients with benign breast tumors. The control group involved 21 healthy people. Our study includes anthropometric, clinical, biochemical, immunological (the reaction of lymphocyte blast transformation to PHA, insulin, antigens cartilage, IGF-1) and morphological methods of investigation. Patients with obesity associated diseases have reduction of proliferative activity in response to PHA and increased frequency of autoimmune reactions to insulin suppressed by prostaglandin-synthesizing cells and/or cells with histamine receptors (144/239 vs. 2/21, p <0,001). This type of cell-mediated immunity is correlated with the anthropometric data (BMI, waist circumference, WC/height) and disorders of carbohydrate metabolism. Conclusions. Despite the differences of diseases associated with visceral obesity, the fact united them is the presence of metabolic disorders, the development of an autoimmune response to insulin, suppressed by prostaglandin synthesizing cells and/or cells with receptors to histamine (more than 60%), which suggests the involvement of the immune system in the genesis of these disorders.

Vancomycin Hypersensitivity Diagnosed by Lymphocyte Blast Transformation

A 15-year-old male admitted for Pott's puffy tumor developed recurrent episodes of fever, diffuse morbilliform rash, eosinophilia, and tubulointerstitial nephritis while on multiple antibiotics. Lymphocyte blast transformation (LBT), a method of detecting cellular immune response by measuring levels of interferon-γ(IFN-γ), was used to diagnose vancomycin hypersensitivity and guide antibiotic selection.

CD69 Expression on CD4+ T Lymphocytes after In Vitro Stimulation with Tuberculin Is an Indicator of Immune Sensitization against Mycobacterium tuberculosis Antigens

ABSTRACT The expression of the CD69 antigen on CD4 T lymphocytes after in vitro stimulation with purified protein derivative (2 tuberculin units) was used to evaluate the tuberculin reactivities of 52 individuals from four experimental groups: Mycobacterium bovis BCG-vaccinated healthy individuals with a negative tuberculin skin test (TST) result (group A), BCG-vaccinated healthy individuals with a positive TST result (group B), patients with active tuberculosis (TB) before treatment (group C), and individuals with clinically inactive TB who had previously completed a prescribed course of chemotherapy (group D). The expression of CD69 on CD4 T lymphocytes was significantly higher in patients with active TB (16.2% ± 7.3%), individuals with clinically inactive TB (10.5% ± 7.4%), and healthy individuals with a positive TST result (15.5% ± 7.2%) than in healthy individuals with a negative TST result (3.8% ± 4.3%) (P < 0.005). We confirmed the correlation between CD69 antigen expression on T lymphocytes after stimulation with tuberculin and the TST induration diameter (Spearman rho = 0.783; P < 0.001), an assay for gamma interferon (the Quantiferon-TB assay; Spearman rho = 0.613; P < 0.001), and the lymphocyte BLAST transformation test (Spearman rho = 0.537; P < 0.001). Our results demonstrate the usefulness of the determination of CD69 on CD4 T lymphocytes after in vitro stimulation with tuberculin as a rapid indicator of immune sensitization against Mycobacterium tuberculosis.