Variant Type X91+ Chronic Granulomatous Disease: Clinical and Molecular Characterization in a Chinese Cohort

Purpose: We aimed to report the clinical and immunological characteristics of variant type X91+ CGD in a Chinese cohort.Methods: The clinical manifestations and immunological phenotypes of X91+ CGD patients were collected. Dihydrorhodamine (DHR) analysis was performed to evaluate neutrophil function. Gp91phox protein expression was determined by flow cytometry-based extracellular staining with the monoclonal antibody (mAb) 7D5. Results: X91+CGD patients accounted for 8% (7/85) of all patients with GCD. The median onset age in the 7 X91+ CGD patients was 4 months. Six patients received the same BCG vaccine strain, and three had probable BCG infections. Moreover, 4 patients were highly suspected of having Mycobacterium tuberculosis infection. Recurrent infections of the lungs and soft tissues (3/7) were the most common symptoms. Two patients had noninfectious recurrent oral ulcers and received interferon gamma (IFN-γ) treatment afterward. In our cohort, the stimulation index (SI) of the 7 X91+ CGD patients ranged widely from 1.9 to 67.5, while the SI ranged from 1.2 to 35.7 in patents with X910 CGD. The level of SI between these two groups was statistically significant (P<0.05). CYBB mutations associated with X91+CGD were usually located in or near the FAD and NADPH binding domains. Three new X91+ CGD related mutations (c.1462-2 A>T, c.1243C>T and c.925G>A) were identified. Conclusions: Variant type X91+ CGD may have varied severities of clinical manifestations. Moreover, the laboratory findings of X91+ CGD could present with a moderate neutrophil stimulation index. We should deepen our understanding of the X91+ variant CGD to prevent missed diagnosis.

Antiviral Therapeutic Potential of Curcumin: An Update

The treatment of viral disease has become a medical challenge because of the increasing incidence and prevalence of human viral pathogens, as well as the lack of viable treatment alternatives, including plant-derived strategies. This review attempts to investigate the trends of research on in vitro antiviral effects of curcumin against different classes of human viral pathogens worldwide. Various electronic databases, including PubMed, Scopus, Web of Science, and Google Scholar were searched for published English articles evaluating the anti-viral activity of curcumin. Data were then extracted and analyzed. The forty-three studies (published from 1993 to 2020) that were identified contain data for 24 different viruses. The 50% cytotoxic concentration (CC50), 50% effective/inhibitory concentration (EC50/IC50), and stimulation index (SI) parameters showed that curcumin had antiviral activity against viruses causing diseases in humans. Data presented in this review highlight the potential antiviral applications of curcumin and open new avenues for further experiments on the clinical applications of curcumin and its derivatives.

Effects of Two Polysaccharides From Traditional Chinese Medicines on Rat Immune Function

The aim is to study the immune function effect of two polysaccharides extracted from traditional Chinese herbs on rats. Ultrasonic-assisted extraction was used to extract the polysaccharide from traditional Chinese medicines. MTT assay was used to determine the effects of two polysaccharides on the conversion of pig peripheral T lymphocytes. For this, 24 Sprague–Dawley rats were selected for the clinical trial and divided into groups B (blank), CK (cyclophosphamide inhibitory control), AP (angelica polysaccharide), and RIP (radix isatidis polysaccharide). Except for group B, other groups can induce the immunodeficiency by using cyclophosphamide. Rats of the AP and RIP groups were given gavage of 1 mL of AP and RIP. The blood was sampled from the eyeball on days 0, 7, 14, 21, 28, and 35, respectively, to determine immune cells, IgG and IgM of immunoglobulin, body weight, and spleen index.Results: The average content of AP and RIP was 51.27 and 14.8%, and the extraction rate was 75.23 and 60.94%. The maximum stimulation index was 1.407 when the concentration of AP was 8,000 μg mL−1 and 1.5 when the concentration of RIP was 125 μg mL−1. Both kinds of polysaccharides can alleviate the decline of white blood cells, lymphocytes, monocytes, neutrophils, and serum IgG and IgM caused by cyclophosphamide. The two polysaccharides can regulate the rapid recovery of weight in immunosuppressed rats and increase the spleen index of immunosuppressed SD rats. The polysaccharides from the two traditional Chinese medicines can alleviate the immunosuppression caused by cyclophosphamide and promote the immune function of the body, which can be used as raw material resources of new veterinary medicine.

First Report of Erythema Multiforme Minor Caused by Raloxifene Hydrochloride

Raloxifene is a drug used in postmenopausal women with osteoporosis. Although hot flashes are known side effects of raloxifene, to the best of our knowledge, erythema multiforme (EM) minor has not been previously reported. Herein, we report about a 74-year-old woman who developed EM minor after the drug alfacalcidol was changed to raloxifene to treat osteoporosis. Skin biopsy revealed a suspicious eczematous drug reaction. The drug-induced lymphocyte stimulation test showed a positive result. The stimulation index was 2.2, and there were no other suspected drugs. Based on these results, we diagnosed the condition as EM minor caused by raloxifene. The patient’s symptoms disappeared after the use of antihistamine drugs and topical steroids. In conclusion, raloxifene can cause EM minor in rare cases.

New opportunities in the diagnosis of asthma with sensitization to Aspergillus spp.

BACKGROUND:Diagnosis of asthma with sensitization toAspergillusspp. is becoming increasingly important due to the severe, uncontrolled course of the disease and the possibility of the formation of allergic bronchopulmonary aspergillosis. AIM:To evaluate the possibility of using the basophil activation test using flow cytometry for the diagnosis of asthma with sensitization toAspergillusspp. MATERIALS AND METHODS:118 patients with asthma were examined. The levels of total IgE and specific IgE to aeroallergens were determined in the blood serum by the enzyme immunoassay. Basophil activation was studied by flow cytometry using the Allergenicity kit (Cellular Analysis of Allergy, Beckman-Coulter, USA). The allergenAspergillus fumigatus(Alcor Bio, Russia) was used to stimulate basophils. RESULTS:The first group consisted of 57 patients with asthma without sensitization toAspergillusspp. The second group included 36 patients with asthma with sensitization toAspergillusspp. The third group consisted of 25 patients with allergic bronchopulmonary aspergillosis. The number of basophils activated by theAspergillusfumigatusallergen in patients with asthma with sensitization toAspergillusspp. and allergic bronchopulmonary aspergillosis was significantly higher than in the asthma group and amounted to 8.1 [5.2; 20.9]% and 84.6 [75.7; 94.0]%, respectively (p 0.001). The stimulation index in the study groups ranged from 0.7 to 72.6. The optimal diagnostic point (cut off) for identifying patients with asthma withAspergillusspp. sensitization there was an stimulation index value of more than 2.4, and for patients with allergic bronchopulmonary aspergillosis 15.95. Among all patients with sensitization toAspergillusspp. a positive correlation was established between the level of specific IgE toAspergillusspp. and the percentage of basophils activated by the allergenAspergillusfumigatus(r= 0.792,p 0.001) and stimulation index (r= 0.796,p 0.05). CONCLUSIONS:The basophil activation test can be used as an additional diagnostic method for asthma with sensitization toAspergillusspp. and allergic bronchopulmonary aspergillosis.

Immune Response of Gamma-Irradiated Inactivated Bivalent Polio Vaccine Prepared plus Trehalose as a Protein Stabilizer in a Mouse Model

<b><i>Introduction:</i></b> Poliovirus causes paralysis by infecting the nervous system. Currently, 2 types of polio vaccine are given in many countries in polio eradication program including inactivated polio vaccine (IPV) and oral polio vaccine (OPV). Because of OPV-related paralysis, OPV should be replaced by IPV. <b><i>Methods:</i></b> The aim of this study was to prepare the gamma-irradiated IPV and determine its effectiveness compared with the commercial vaccine (OPV) in the mouse model. The virus titration of OPV was determined and then inactivated by the appropriate dose of gamma radiation into an irradiated vaccine formula. The vaccine was inoculated in BALB/c mice in 2 different formulations of intramuscular injection with 2-week intervals. The level of anti-polio-neutralizing antibody and polio-specific splenocyte proliferation assay were evaluated by collecting the blood samples and spleens of the vaccinated groups with conventional vaccine and irradiated vaccine. <b><i>Results:</i></b> There was a significant increase in the neutralizing antibody titration between all of the vaccinated groups and negative control group (A) (<i>p</i> &#x3c; 0.05). And it shows that the IPV by gamma irradiation has the highest antibody titration. Also, the increasing of stimulation index value in the B* group, F group, and G group was the most against other groups. Furthermore, the neutralizing anti-serum titer and splenic lymphocyte proliferation assay show humoral and cellular immunity were significantly increased in the irradiated vaccine group as compared with conventional group. <b><i>Conclusion:</i></b> According to the results, gamma-irradiated IPV could induce humoral and cellular immunity in vaccinated mouse groups, so the irradiated poliovirus could be recommended as a good candidate vaccine to prevent the transport of poliovirus to the central nervous system and thus protect against paralysis.

Immunomodulatory and Antioxidant Activities of Select Indonesian Vegetables, Herbs, and Spices on Human Lymphocytes

Edible plants have attracted increasing attention as functional foods as they are rich in bioactive compounds with health benefits, including antioxidant and immunomodulatory activities. However, scientific evidence of these health effects is limited. This study is aimed at determining antioxidant and immunomodulatory activities of 25 select vegetables, herbs, and spices commonly consumed in Indonesia. Phytochemical profiles were determined by measuring total flavonoid content and 1H-NMR. Human blood lymphocyte cells were used to probe the immunomodulatory potency and treated with the methanol extract of these vegetables, herbs, and spices. The results showed the enhanced propensity for all tested plant extracts to stimulate lymphocyte proliferation, except Pandanus amaryllifolius. Etlingera elatior, Ocimum xcitriodorum, Kaempferia galanga, and Apium graveolens had the highest lymphocyte cell proliferation stimulation index (SI) at concentrations of 41.67, 16.67, 4.17, and 2.5 mg/mL culture, respectively (SI 2.21 ± 0.05 , 2.62 ± 0.12 , 3 ± 0.05 , and 2.64 ± 0.07 , respectively). The NMR spectra of these four most potent plants showed low peaks in the aromatic/phenolic area and several other peaks indicating the presence of terpenoid, steroid, amino acid, and sugar compounds. The results demonstrate the immunomodulatory potential of all vegetables, herbs, and spices, except P. amaryllifolius, although this potential did not necessarily correlate with flavonoid content and antioxidant activity. Nevertheless, this research showed promising health effect, particularly immunomodulation, of the various local plants. Further elaboration on the specific immunomodulatory activity will be interesting.

Dynamics of basophil reactivity in the patients with allergic rhinitis on the background of allergen-specific immunotherapy

BACKGROUND: Allergen-specific immunotherapy (ASIT) helps to reduce spontaneous and to induce basophil reactivity, which is one of the major mechanisms of its effectiveness. A number of studies, though, have not confirmed the informative value of the basophil activation test (BAT), in case of ASIT, with house dust mites, meadow grass pollen, and wasp venom. BAT outcomes are likely to be influenced by the properties of the sensitizing allergen, the duration of ASIT, and the BAT markers that are used. AIMS: To evaluate the basophils reactivity in the patients sensitized to weed pollen and treated with allergen-specific therapy. MATERIALS AND METHODS: The study involved 60 patients (aged 1655) with allergic rhinitis sensitized to weed pollen. The diagnosis was based on the skin test outcomes as well as on identifying specific antibodies to major allergens of ragweed, wormwood, timothy, and birch. The pre-seasonal ASIT involved water-salted extracts of pollen allergens during 2 years. Clinical symptoms and the immunotherapy effectiveness were evaluated 6 and 18 months after the ASIT started. BATs were made before the treatment, after the first and second courses of pre-seasonal ASIT, which corresponded to zero, the 3rd and the 15th months of the treatment. The activated basophils were identified based on the CRTH2pos CD203brightCD3neg phenotype. RESULTS: The first treatment course was completed by 52 patients, while the second one by 43 patients. A statistically significant decrease in spontaneous basophil reactivity was detected only after the second course of ASIT. The decrease in the allergen-induced basophil activation was identified in 67.4% of the samples obtained after the first ASIT course, and in 67.6% of samples taken after the second immunotherapy course. A decrease in the BAT stimulation index (BAT SI) was identified mainly in the patients featuring positive treatment outcomes. CONCLUSIONS: ASIT influences on the basophil reactivity indicators in the patients sensitized to weed pollen. The basophil activation test may be considered as a potential predictor of ASIT effectiveness.

IL-2 and IFN-[gamma] are biomarkers of SARS-CoV-2 specific cellular response in whole blood stimulation assays

Objectives: A proper description of the immune response to SARS-CoV-2 will be critical for the assessment of protection elicited after both infection and vaccination. Uncoupled T and B cell responses have been described in acute and convalescent patients and exposed individuals. We aimed to assess the potential usefulness of whole blood stimulation assays to identify functional cellular immune responses to SARS-CoV-2. Methods: Blood from COVID-19 recovered individuals (5 months after infection) and negative subjects was stimulated for 24 hours with HLA predicted peptide megapools of the Spike and Nucleoprotein, or the mixture of them. After stimulation, cytokines were quantified using a beads-based multiplex assay. Results: Interleukin-2 and IFN-γ were found to be specific biomarkers of SARS-CoV-2 cellular response. Using the Spike and Nucleoprotein mixture, 91.3% of COVID-19 recovered individuals presented an IL-2 stimulation index over the cut-off, while 82.6% showed IFN-γ. All the negative individuals presented an IL-2 response under the cut-off, while 5.3% of these subjects presented positive IFN-γ stimulation indexes. Moreover, IL-2 production correlated with IgG levels for Spike 1, RBD, and Nucleocapsid. Conclusion: We demonstrate the potential of whole blood stimulation assays and the quantification of IL-2 and IFN-γ for the analysis of SARS-CoV-2 functional cellular responses.

Clinical and immunological characteristics of severe bronchial asthma with Aspergillus sensitization

Aim: to assess the clinical and immunological characteristics of patients with severe bronchial asthma (BA) with Aspergillus sensitization and to study the possibilities of applying basophil activation test (BAT) using flow cytometry to detect mycogenic sensitization. Patients and Methods: 93 patients with severe BA were examined. Therapy had 4 or 5 steps according to GINA 2019 recommendations. TARC (thymus and activation-regulated chemokine) content, the number of eosinophils, the levels of total IgE and specific IgE to 10 allergens were determined in blood serum by enzyme immunoassay. BAT with Aspergillus fumigatus allergen was performed in vitro using flow cytometry. Results: Aspergillus sensitization was detected in 33 (35.4%) patients with severe BA. In patients with BA and Aspergillus sensitization, the disease course was uncontrolled, and patients in this group were credibly more likely to receive oral glucocorticosteroids. The number of basophils (activated by the A. fumigatus allergen) and the stimulation index in patients with BA and Aspergillus sensitization were significantly higher than in patients with BA (9.9 (6.0–24.0) % vs. 3.6 (2.0–5.4) %; (p=0.000) and 4.25 (2.49–9.30) vs. 0.94 (0.75–1.16); (p=0.000)). Significant differences in TARC content were obtained in the groups of patients with severe BA and Aspergillus sensitization and patients with BA (625.0 (418.4–875.0) pg/mg versus 406.0 (210.0–561.0) pg/mg; p=0.001). A negative correlation was determined between TARC levels and a decrease in forced expiratory volume in 1 second (FEV1) (r=-0.70, p<0.05), and between positive correlation and absolute eosinophil count (r=0.81, p<0.05) and level of specific IgE to Aspergillus (r=0.36, p<0.05). Conclusion: Aspergillus sensitization is associated with an uncontrolled BA course. An additional method for diagnosing mycogenic sensitization is the BAT. The TARC concentration can serve as a biomarker of an active inflammatory response. KEYWORDS: Aspergillus spp., severe bronchial asthma, mycogenic sensitization, basophil activation test, TARC. FOR CITATION: Kozlova Ya.I., Frolova E.V., Uchevatkina A.E. et al. Clinical and immunological characteristics of severe bronchial asthma with Aspergillus sensitization. Russian Medical Inquiry. 2021;5(1):10–16. DOI: 10.32364/2587-6821-2021-5-1-10-16.