Utilización precoz de tocilizumab en pacientes internados con COVID-19 grave y crítica: un estudio multicéntrico en la Provincia de Buenos Aires / Early use of tocilizumab in hospitalized patients with severe and critical COVID-19 in the Province of Buenos Aires: a multicentric study

Resumen Introducción: El objetivo principal del estudio fue evaluar la mortalidad en los pacientes con COVID-19 graves y críticos, que recibieron tocilizumab (TCZ) -un antagonista monoclonal del receptor de IL-6- de forma temprana vs. tardía. Métodos: Cohorte retrospectiva multicéntrica de pacientes >18 años internados con COVID-19 desde el 1/7/2021-1/8/2022, con 5-7 puntos de gravedad inicial (GI) según Escala de la OMS. Se consideró adminis tración temprana o tardía a la infusión de TCZ ≤ ó > a 48 h del ingreso. Las variables de resultado fueron mortalidad a 28 días y cambio de la GI. Los factores relacionados con la mortalidad fueron evaluados con regresión de Cox. Resultados: Se incluyeron 266 pacientes, 159(60%) varones; edad 58(± 15); con hipertensión arterial (43%), obesidad (37%) y diabetes (27%);70 presentaban GI = 5 (oxígeno suplementario), 143 GI = 6 (ventilación no inva siva o cánula nasal de alto flujo) y 53 GI = 7 (ventilación mecánica invasiva). La mortalidad a 28 días fue 42%, asociada independientemente a: edad, obesidad, GI, días entre la internación y administración del TCZ, y días entre la fecha de inicio de síntomas y el TCZ. La mortalidad para GI 5, 6 y 7 fue 26%, 39% y 72%, respectivamente; 76% y 62% de los pacientes permanecieron estables o mejoraron la GI a los días 3 y 7 de la infusión de TCZ. La mortalidad a 28 días fue 39% (TCZ temprano) vs. 57% (TCZ tardío); p = 0.02; HR = 0.63[0.41-0.99, p = 0.05]). Discusión: Estos resultados apoyan la administración temprana de TCZ en pacientes con COVID-19 grave y crítica.

Abstract Introduction: Tocilizumab (TCZ), an IL-6 receptor antagonist monoclonal antibody is warranted in severe and critically-ill COVID-19 patients. The objective was to evaluate 28-day mortality of patients with severe or critical COVID-19 treated with early vs delayed TCZ. Methods: Multicenter, retrospective cohort study in cluding patients>18 years hospitalized between 7/1/2021- 8/1/2022 with confirmed COVID-19, with 5, 6 and 7 points of WHO Ordinal Initial Severity Scale [SS]. Early or late administration was considered if TCZ was administered before or after 48 hours from admission. Outcomes were28-day mortality and change of SS. Factors related to 28-day mortality were evaluated with Cox regression. Results: 266 patients were included, 159(60%) male; aged 58(± 15); frequent comorbidities were hypertension (42%), obesity (37%) and diabetes (27%). Seventy patients had a SS = 5 (Supplemental O2), 143 had SS = 6 (NIV/ HFNC), and 53 had SS = 7 (IMV). 28-day mortality was 42%(112/266); predictors were age, obesity, higher SS, days between hospitalization and TCZ administration, and fewer days between symptoms onset and TCZ. Mortality of SS 5, 6 and 7 was 26%, 39% and 72% respectively. Com pared with baseline SS points, 76% and 62% of patients remained stable or improved on days 3 and 7 since TCZ administration. 28-day mortality was lower when TCZ was administered before 48 hours (39% vs 57%; p = 0.02; HR = 0.63;[0.41-0.99, p = 0.05]). Discussion: This study supports the early use of TCZ in patients with severe or critical COVID-19.

Increased expression of interleukin-6 gene in gastritis and gastric cancer

Helicobacter pylori (H. pylori) induces an intense inflammatory response, mediated by proinflammatory cytokines, including interleukin (IL)-6 and its membrane receptor (IL-6R), which activates important signaling pathways in the development of gastric disease and cancer. We investigated the gene and protein expression of IL-6 and IL-6R and the influence of polymorphisms rs1800795, rs1800796, and rs1800797 on its gene expression together with H. pylori infection. Furthermore, an in-silico analysis was performed to support our results. Gastric biopsies were obtained from patients with gastric symptoms and patients with gastric cancer (GC) and were divided into groups (Control, Gastritis, and Cancer). H. pylori was detected by PCR. Real-time-qPCR was employed to determine gene expression, and western blot assay was used to analyze protein expression levels. PCR-RFLP was used to characterize IL-6 polymorphisms. Bioinformatics analyses were performed using the Gene Expression Omnibus (GEO) database and GEO2R to screen out differentially expressed genes (DEGs). H. pylori was detected in 43.3% of the samples. Statistically significant differences were found for IL-6 (P=0.0001) and IL-6R (P=0.0005) genes among the three groups, regardless of the presence of H. pylori. Among patients with H. pylori infection, the IL-6 and IL-6R gene and protein expressions were significantly increased, highlighting IL-6 gene overexpression in patients with GC. No statistically significant differences were found for the rs1800795, rs1800796, and rs1800797 polymorphisms compared to IL-6 gene expression. The results indicated that the IL-6 polymorphisms do not influence its expression, but IL-6 and IL-6R expression seems to be altered by the presence of H. pylori.

The change of interleukin -6, soluble interleukin - 6 receptor and soluble glycoprotein 130 levels in patients with lung injury caused by paraquat poisoning and its clinical significance / 中国医师进修杂志

Objective:To investigate the change of interleukin-6(IL-6), soluble interleukin-6 receptor(sIL-6R) and soluble glycoprotein 130(sgp130) levels in patients with lung injury caused by paraquat poisoning and its clinical significance.Methods:Fifty patients with paraquat poisoning and lung injury admitted to Ba′nan District People′s Hospital of Chongqing from December 2017 to December 2019 were selected as the observation group, and 30 healthy patients were selected as the control group. The serum levels of IL-6, sIL-6R and sgp130 in two groups was detected with double-antibody sandwich enzyme-linked immunosorbent assay and compared. Pearson correlation analysis was performed on the relationship between serum IL-6, sIL-6R and sgp130 and the acute physiological and chronic health status assessment(APACHE)- Ⅱ score of patients with lung injury caused by paraquat poisoning. The receiver operating characteristics (ROC) curve was used to analyze the diagnostic value of serum IL-6, sIL-6R and sgp130 on paraquat poisoning-induced lung injury.Results:The serum levels of IL-6, sIL-6R, and sgp130 in the observation group were significantly higher than those in the control group: (108.62 ± 11.39) ng/L vs. (57.41 ± 7.63) ng/L, (73.28 ± 6.94) ng/L vs. (45.13 ± 6.57) ng/L, (435.64 ± 36.52) mg/L vs. (281.71 ± 68.35) mg/L, and the differences were statistically significant ( P<0.05). The results of correlation analysis showed that the serum levels of IL-6, sIL-6R, and sgp130 in the observation group were significantly positively correlated with the APACHE-Ⅱ score ( r = 0.824, 0.937, 1.215, P<0.05). ROC curve analysis results showed that when serum IL-6, sIL-6R and sgp130 had the best diagnostic cut-off values of 83.96 ng/L, 68.51 ng/L and 367.42 mg/L respectively for the lung injury caused by paraquat poisoning, the sensitivity was 84.80%, 78.20% and 87.10%, and the specificity was 70.30%, 85.50% and 81.00%. Conclusions:The expression levels of IL-6, sIL-6R and sgp130 in serum are closely related to the severity of lung injury caused by paraquat poisoning, and IL-6, sIL-6R and sgp130 have good diagnostic value in lung injury caused by paraquat poisoning.

The effect of targeting interleukin-6 receptor on leukocyte production after acute myocardial infarction / 实用医学杂志

Objective To observe the effects of Tocilizumab on white blood cell (WBC) after acute myocardial infarction (MI) and explore its potential to treat MI.Methods Rats were divided into 3 groups:control,MI,and MI treated.Serum from individual mouse was collected before and after subcutaneously Tocilizumab treatment.The level of interleukin-6 (IL-6),the number of WBC and the ratio of active hematopoietic stem cell (HSC)was tested by ELISA,flow cytometry and blood routine examination.The fibrosis of heart tissue was observed by immunohistochemistry.Results The IL-6 level and the number of the WBC were reduced after Tocilizumab treatment.It indicates the effect of inhibiting the activity of HSC and improving the situation of cardiocytes remodeling.Conclusion Tocilizumab could inhibit the generation of WBC and re-construct myocardium after MI.

Interleukin-6 Receptor Polymorphisms Contribute to the Neurological Status of Korean Patients with Ischemic Stroke

To investigate the contribution of the interleukin-6 receptor (IL-6R) gene single nucleotide polymorphisms (SNPs) to the neurological status of Korean patients with ischemic stroke (IS), two SNPs of the IL-6R gene (rs4845617, 5 UTR; rs2228144, Ala31Ala) were selected. IS patients were classified into clinical phenotypes according to two well-defined scores: the National Institutes of Health Stroke Survey (NIHSS) and the Modified Barthel Index scores. There were 121 IS patients and 291 control subjects. The SNP rs4845617 significantly contributed to the neurological status of patients with IS (P = 0.011 in codominant model 2, P = 0.006 in recessive model, and P = 0.008 in log-additive model). Allele frequencies of rs4845617 and rs2228144 demonstrated no significant difference in IS patients and controls. The AG and GG haplotypes differed between the NIHSS 1 (NIHSS scores or = 6) group in patients with IS (P = 0.014, P = 0.0024). These results suggest that rs4845617 of the IL-6R gene is associated with the neurologic status of Korean patients with IS.

The expression of IL-6Ralpha and Gp130 in fallopian tubes bearing an ectopic pregnancy / 대한해부학회지

Women with tubal ectopic pregnancies have high levels of circulating interleukin 6 (IL-6). IL-6 treatment in vitro significantly reduces the ciliary activity of tubal epithelium. The effects of IL-6 on target cells occur via the formation of a high-affinity complex with its receptors IL-6Ralpha and glycoprotein 130 (Gp130). IL-6Ralpha is specifically expressed in the cilia of the epithelial cells. In this study, we performed a quantitative reverse transcriptase polymerase chain reaction to determine the mRNA expression of IL-6Ralpha and Gp130 in the fallopian tubes obtained from 12 women with ectopic pregnancies, 12 women with normal pregnancies, and 12 healthy nonpregnant women in the luteal phase of their menstrual cycle. Fallopian tubes were evaluated from specimens taken during tubal ligation in normal pregnancies and nonpregnant fertile women or during tubal surgery in ectopic pregnancies. We observed that IL-6Ralpha mRNA expression in fallopian tubes was increased in ectopic pregnancy compared with that in the midluteal phase. We also found that the Gp130 mRNA expression was significantly lower in fallopian tubes from ectopic pregnancies than in those from nonpregnant women during the midluteal phase of their menstrual cycle, although its expression was noticeably high in fallopian tubes in the midluteal phase, which suggests that high Gp130 levels may possibly contribute to embryo transport into the uterus.

Association of single nucleotide polymorphism of interleukin 6 receptor gene and metabolic syndrome / 中华内分泌代谢杂志

The association of interleukin 6 receptor ( IL-6R ) gene - 183 A/G ( rs4845617 ) and Asp358 Ala (rs8192284 A/C) polymorphisms with metabolic syndrome was investigated in Chinese Han population.The result showed that the frequencies of AA genotype and A allele were higher in patients with metabolic syndrome ( MS ) than those in healthy subjects ( P＜0.05 or P＜0.01 ).The risk of MS in patients with A allele was 1.643 folds of that with allele C(95％ CI 1.163-2.320,P＜0.01 ).No differences were found in the genotype and allele frequencies of -183A/G between two groups ( P＞0.05 ).The Asp358Ala polymorphism of IL-6R was significantly associated with MS in Chinese Han population.

Association of interleukin-6 receptor gene polymorphism with obesity / 中华内分泌代谢杂志

To examine the relationship between genetic variants in the interleukin-6 receptor gene and obesity. The result showed that the carriers of Asp358 homozygotes had higher risk in developing obesity when compared with Ala358 homozygotes (OR = 1.32,95% CI 1.07-1.68, P = 0. 041). Interleukin-6 receptor gene pulymorphism was significantly associated with obesity.

Density and Affinity of IL-6 Receptors in Human Leukemic Cells / 中国肿瘤生物治疗杂志

Objective: To make a study of density and affinity of IL-6R in human leukemic cell lines, and discuss the affection of high affinity IL-6R to the targeted treatment of leukemia with IL-6-PE40 fusion protein. Methods: Radial binding assay with scatchard plot and FACS were used to analysis the density and affinity of IL-6R and protein expression of IL-6Rα and β subunits in totally 8 representative human leukemic cell lines. Results: Myelocytie, monocytic and erythrocytic leukemic cell lines U937, HL-60, KG1 and TF1 express high affinity IL-6R, whose average density per cell is 2 502,2 874, 2 319 and 9 329 respectively, however no 125I-IL-6 binding was detected on chronic myelocytic leukemic cell line K562 and lymphoblastic leukemic cell lines such as Raji, CEM and HUT28. These results correlate with those of FACS highly. Conclusion:These observations suggest that acute nonlymphoblastic leukemic cells may be more suitable for targeted treatment with IL-6-PFA0 fusion protein.

The role of interleukin 6 and interleukin 6 receptor in gliomas / 中国癌症杂志

Purpose:To discuss the role of interleukin 6 (IL 6) and IL 6 receptor (IL 6R) in gliomas. Methods:We detected IL 6 and IL 6R gene in 30 human glioma tissue specimens and adult normal astrocytes. A complementary DNA copy of total RNA was synthesized and amplified with specific primers using the reverse transcription polymerase chain reaction (RT PCR) method. Results:IL 6 gene was positive of in 24 (80.0%) samples of gliomas, while IL 6R gene was positive of in 26 (86.7%) cases. Co expression of IL 6 and IL 6R was identified in 22 (73.3%) specimens of gliomas. On the other hand,the expression of IL 6 was weakly positive in adult normal astrocyte, but IL 6R was negative.Conclusions:Our results indicate that IL 6/IL 6R autocrine or paracrine loop may exist in glioma, which could enhance the glioma cell proliferation.

Activation of Stat Protein in Synovial Cell in Rheumatoid Arthritis / 대한류마티스학회지

OBJECTIVE: To characterize Stat activity in synovial tissue in rheumatoid arthritis (RA) in order to see if Stat molecule contributes to the pathogenesis of RA by regulatory expression of genes that play an important role in inflammation and tissue destruction. METHODS: Synovial tissue were obtained immediately after operative excision. Immuno-histochemistry was done with the antibodies for Stat 3 and Stat 5. Cells were stimulated with interleukin 6 (IL-6) and soluble interleukin 6 receptor (sIL-6R) or steroid using chambered slide. In supershift experiment, cell extracts were incubated with 0.5ng of 32P-labelled double-stranded oligonucleotide probe. Samples were resolved on 4.5% polyacrylamide gels, which was transferred to polyvinylidene fluoride membranes. Anti-phosphotyrosine Stat 3 antibody was used for Western blotting. RESULTS: Stat 3 was not shown on the synovial tissue section done by immuno-histochemistry. However, activated Stat 3 was expressed on cultured synovial cell stimulated with IL-6 and sIL-6R, and also with IL-6 and dexamethasone using chambered slide. In contrast to Stat 3, activated Stat 5 was expressed on the synovial tissue section, especially around blood vessel. CONCLUSION: Stat is activated in cultured synovial cells as shown in other immune associated cells, and IL-6 is the strong activator of Stat 3. Further analysis of the regulation of Stats in synovitis and the role of Stats in driving synovial inflammation will yield insight into the pathogenesis of RA and the development of novel therapeutic modality.

Interleukin-6 and Soluble Interleukin-6 Receptor in Degenerative Arthrits / 대한류마티스학회지

Unlike other soluble receptors, the soluble interleukin-6 receptor (sIL-6R) cooperates with IL-6 to activate gp130 of effector cell. As the IL-6 and sIL-6R are important in the rheumatoid disease, this study was designed to measure concentration of IL-6 and sIL-6R in synovium and synovial fluid of the degenerative arthritis. The synovium and synovial fluid were obtained during total knee replacement arthroplasty. The synovium was taken from eleven patients, and synovial fluid taken from sixteen patients. Same patients between two groups were seven. Tissue cultures of the synovial tissues were done with 10% FBS for 72 hours. After irrigation, thery were incubated for 48 hours without FBS, and the culture media and the synovial fluid were collected after centrifuged at 2500rpm for 10 minutes. The level of IL-6 and sIL-6R were measured by quantitative sandwich enzyme immunoassay technique. RESULTS: In the synovium, the IL-6 level was 5.1+/-0.12ng/ml, and the sIL-6R level was 0.41+/-0.25ng/ml. In the synovial fluid, the IL-6 level was 0.09+/- 0.15ng/ml, and the sIL-6R level was 10.37+/-3.28ng/ml. These results show that IL-6 concentration was measured highly in two groups, especially in synovium (sixty times), and the sIL-6R concentration was measured significantly high in synovial fluid (twenty-five times). CONCLUSION: The IL-6 and sIL-6R were elevated in degenerative arthrits. We confirmed the source of IL-6 was synovium (very high in synovial tissue culture media), but we need further study for the source of sIL-6R as it was remarkably elevated as IL-6 and its level was lower than serum.