Molecular analysis of alpha‐ and beta‐thalassemia in Meizhou region and comparison of gene mutation spectrum with different regions of southern China

BackgroundThalassemia is a group of inherited autosomal recessive hemolytic anemia disease caused by reduced or absent synthesis of globin chain/chains of hemoglobin. Only few studies showed the molecular characterization of α‐ and β‐thalassemia in Meizhou city of China.MethodsA total of 22,401 individuals were collected; hematological and hemoglobin electrophoresis analysis and thalassemia genetic testing were performed.ResultsEleven thousand and thirty (49.24%) cases with microcytosis (mean corpuscular volume (MCV) < 82 fl), 11,074 (49.44%) cases with hypochromia (mean corpuscular Hb (MCH) < 27 pg) in 22,401 subjects, 11,085 cases with abnormal hemoglobin results were identified in subjects aged ≥6 months. 7,322 (32.69%) subjects harbored thalassemia mutations, including 4,841 (21.61%) subjects with α‐thalassemia, 2,237 (9.99%) with β‐thalassemia, and 244 (1.09%) with α‐thalassemia combined β‐thalassemia. 18 genotypes of α‐thalassemia mutations and 27 genotypes of β‐thalassemia mutations were characterized. The most frequent α gene mutation was ‐‐^SEA (64.69%), followed by ‐α^3.7 (19.93%), ‐α^4.2 (7.73%), α^CSα (3.97%), and α^WSα (2.83%). The six most common β‐thalassemia mutations were IVS‐II‐654 (C>T) (39.79%), CD41‐42 (‐TCTT) (33.02%), −28 (A>G) (10.38%), CD17 (A>T) (9.08%), CD27‐28 (+C) (2.14%), and CD26 (G>A) (2.02%). In addition, MCV and MCH were sensitive markers for α‐ and β‐thalassemia except for ‐α^3.7/αα, ‐α^4.2/αα, α^CSα/αα, α^WSα/αα, and β^Cap+40−43/β^N.ConclusionsThe ‐‐^SEA, ‐α^3.7, and ‐α^4.2 deletions were the main mutations of α‐thalassemia, while IVS‐II‐654 (C>T), CD41‐42 (‐TCTT), −28 (A>G), and CD17 (A>T) mutations of β‐thalassemia in Meizhou. There were some differences in thalassemia mutation frequencies in Meizhou city from other populations in China.     

Prenatal diagnosis of thalassemia in 695 pedigrees from southeastern China: a 10‐year follow‐up study

Thalassaemia is highly prevalent in southeastern China. This 10‐year follow‐up study aimed to characterize the genotype and karyotype of thalassaemia in fetal samples derived from thalassemia carriers in Fujian province, southeastern China. A total of 476 prenatal samples from 472 couples carrying α‐thalassaemia traits and 224 samples from 223 couples carrying β‐thalassaemia traits were collected for STR analysis, detection of thalassemia genotypes and karyotyping. The common deletional α‐thalassemias and rare thalassemia genotypes were detected using Gap‐PCR assay, and the common β‐globin gene mutations were detected using PCR‐RDB assay. We detected 43.49% prevalence of α‐thalassaemia minor, 26.05% prevalence of α‐thalassaemia intermediate and major and 1.89% prevalence of rare form among the 476 prenatal samples from couples with α‐thalassaemia, and 85 fetuses with β‐thalassemia heterozygote, 16 with homozygote and 21 with double heterozygote, and a rare β^{IVS−2−654(C→T)}/Chinese G_γ (^Aγδβ)⁰ genotype among the 224 prenatal samples from couples with β‐thalassemia. Karyotyping showed 7 fetuses with abnormal karyotypes. Totally 153 pregnancies were terminated, and genetic diagnosis of thalassemia using fetal umbilical cord blood following induction of labor showed consistent results with prenatal diagnosis. No thalassemia phenotypes were identified in normal infants half a year after birth, and the infants with α‐thalassemia and β‐thalassemia minor had no or mild anemia symptoms, but normal development, while 15 babies with hemoglobin H disease presented moderate anemia symptoms. Our data suggest the pregestational screening of thalassemia, notably compound and rare forms of thalassemia, for couples carrying thalassemia traits.     

Evaluation of intervention strategy of thalassemia for couples of childbearing ages in Centre of Southern China

BackgroundTo describe the free intervention strategy of thalassemia for childbearing couples in Guangzhou.MethodsRoutine hematology examinations were conducted for 137,222 couples. Among them, 37,501 couples who had mean corpuscular volume (MCV) <82 fL or mean corpuscular hemoglobin <27 pg were elected for Hb analysis and the deletions of four common α‐thalassemia mutation. Reverse dot blot for common nondeletional α‐thalassemia and β‐thalassemia was selectively used. Three thousand twenty‐two couples randomly selected were offered all those tests as a control group. Sanger sequencing, multiplex ligation‐dependent probe amplification and next‐generation sequencing were used for rare thalassemia. High‐risk couples were offered prenatal diagnosis at 10–13 weeks’ gestation based on informed consent.ResultsThe carrier rates of α‐, β‐, and αβ‐thalassemia and δβ thalassemia/deletional HPFH were 7.7%, 3.02%, 0.5% and 0.059% respectively. Of them, 1.37% were identified as at‐risk couples and 345 couples terminated the pregnancy. No severe α‐ and β‐thalassemia births were observed. In the control group, two β‐ thalassemia carriers and one case with −α^3.7/αα^QS were misdiagnosed, but all at‐risk couples were found, and we could save 1,523,774 ¥ using our strategy. The cut‐off points of 73.46 fL and 23.25 pg would be useful to find −α⁺/α^T thalassemia.ConclusionThe intervention strategy was cost‐effective and offered reference in population thalassemia screening.     

Association of alpha‐thalassemia and Glucose‐6‐Phosphate Dehydrogenase deficiency with transcranial Doppler ultrasonography in Nigerian children with sickle cell anemia

BackgroundStroke is a devastating complication of sickle cell anemia (SCA) and can be predicted through abnormally high cerebral blood flow velocity using transcranial Doppler Ultrasonography (TCD). The evidence on the role of alpha‐thalassemia and glucose‐6‐phosphate dehydrogenase (G6PD) deficiency in the development of stroke in children with SCA is conflicting. Thus, this study investigated the association of alpha‐thalassemia and G6PD(A⁻) variant with abnormal TCD velocities among Nigerian children with SCA.MethodsOne hundred and forty‐one children with SCA were recruited: 72 children presented with normal TCD (defined as the time‐averaged mean of the maximum velocity: < 170 cm/s) and 69 children with abnormal TCD (TAMMV ≥ 200 cm/s). Alpha‐thalassemia (the α‐3.7 globin gene deletion) was determined by multiplex gap‐PCR, while G6PD polymorphisms (202G > A and 376A > G) were genotyped using restriction fragment length polymorphism—polymerase chain reaction.ResultsThe frequency of α‐thalassemia trait in the children with normal TCD was higher than those with abnormal TCD: 38/72 (52.8%) [α‐/ α α: 41.7%, α ‐/ α ‐: 11.1%] versus 21/69 (30.4%) [α‐/ α α: 27.5%, α ‐/ α ‐: 2.9%], and the odds of abnormal TCD were reduced in the presence of the α‐thalassemia trait [Odds Ratio: 0.39, 95% confidence interval: 0.20–0.78, p = 0.007]. However, the frequencies of G6PDA⁻ variant in children with abnormal and normal TCD were similar (11.6% vs. 15.3%, p = 0.522).ConclusionOur study reveals the protective role of α‐thalassemia against the risk of abnormal TCD in Nigerian children with SCA.     

Molecular epidemiological and hematological profile of thalassemia in the Dongguan Region of Guangdong Province,Southern China

BackgroundThalassemia is a common inherited hematological disease in tropical and subtropical regions. This study aimed to investigate the mutation spectrum of thalassemia in the Dongguan region of southern China and comprehensively analyze hematologic features of thalassemia carriers with various types of globin mutations.MethodsA hematological screening including hematological indices such as mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), and mean corpuscular hemoglobin concentration (MCHC) was conducted in 19 442 people from Dongguan region, Guangdong province of China. Then, 4891 suspected thalassemia carriers were further investigated by genetic analysis of combined NGS and gap‐PCR.ResultsTotally, 2319 (11.9%) cases were diagnosed as carriers of thalassemia, of which 1483 cases (7.6%) were α‐thalassemia, 741 cases (3.8%) were β‐thalassemia, and 95 cases (0.5%) were co‐inheritance of α‐ and β‐thalassemia. In α‐thalassemia carriers, the phenotypic severity increases with the number of nonfunctional α‐globin genes. The patients with –^SEA/α^WSα genotype have less severe clinical phenotypes than those with other Hb H diseases. As for β‐thalassemia, the MCV and MCH in both β⁰ and β⁺ carriers are markedly reduced.ConclusionsThis is the first comprehensive molecular epidemiological survey and hematological profiling of thalassemia in Dongguan area. This study will be benefit for genetic counseling in the clinic and may help pediatricians to make a correct diagnosis of different types of thalassemia.     

Genetic research and clinical analysis of β‐globin gene cluster deletions in the Chinese population of Fujian province: A 14‐year single‐center experience

BackgroundHeterozygotes of HPFH and δβ thalassemia are clinically asymptomatic or have mild hemoglobin (Hb) values. However, when both HPFH and δβ‐thalassemia are coinherited with heterozygous β‐thalassemia, patients may progress to a clinical phenotype of thalassemia intermedia or thalassemia major. The purpose of this study was to characterize the genotypes and analyze the phenotypes of these disorders in Fujian Province, to offer advice for genetic counseling and accurate prenatal diagnosis in this region. A total of 55 001 subjects were participated in thalassemia screening. 142 subjects with HbF levels ≥10%, before the blood transfusion, were selected for further investigation.MethodsMultiplex ligation‐dependent probe amplification (MLPA) and Gap‐PCR were used to screen for three β‐globin gene cluster deletions: Chinese ^Gγ(^Aγδβ)⁰ thalassemia and Southeast Asia HPFH (SEA‐HPFH) deletion and 1357 bp deletion (NG‐000007.3:g.69997‐71353 del 1357).ResultsA total of 142 patients with HbF (≥10%) were enrolled to characterize the molecular basis of β‐globin gene cluster deletions in our study; 22 cases 0.04% (22/55 001) were definitively diagnosed with β‐globin gene cluster deletions. Ten cases were heterozygous for the Chinese ^Gγ(^Aγδβ)⁰‐thal mutations, 10 cases were heterozygous for SEA‐HPFH, and one case was compound heterozygous for SEA‐HPFH and the α‐thal mutation. The 1357 bp deletion (NG‐000007.3:g.69997‐71353 del 1357) was detected in one case. Moreover, the hemoglobin A₂ levels in patients who were heterozygous for Chinese ^Gγ(^Aγδβ)⁰‐thal were statistically lower than in cases with SEA‐HPFH deletion(p < 0.05).ConclusionIn Fujian Province, the prevalence of common β‐globin gene cluster deletions was 0.04%. What''s more, the most common β‐globin cluster deletions are the Chinese ^Gγ(^Aγδβ)⁰ and SEA‐HPFH.     

Antiphospholipid antibodies in autoimmune thyroid diseases

BackgroundAntiphospholipid (aPL) antibodies have been reported in several autoimmune diseases. The aim of this study was to evaluate the frequency of aPL (anti‐cardiolipin antibodies (aCL) and anti‐β2 glycoprotein I antibodies (aβ2GPI)) in patients with autoimmune thyroid diseases (AITD).MethodsOne hundred and ninety‐five patients with AITD (139 Hashimoto''s thyroiditis (HT) patients and 56 Graves'' disease (GD) patients) and 90 healthy blood donors (HBD) were studied. IgG, IgA and IgM aCL and aβ2GPI were determined by ELISA.ResultsOne hundred fifty‐four AITD patients were women and 41 were men. Fifty‐six healthy subjects were women and 34 were men. The median age of patients and the control group was 45 and 38.5 years, respectively. The frequency of aPL was significantly higher in patients with AITD and in patients with HT than in HBD (33.3% vs 11.1%, p < 10⁻³ and 38.1% vs 11.1%, p < 10⁻³). The frequency of aPL in GD was significantly lower than in HT (21.4% vs 38.1%, p = 0.025). In patients with HT, aβ2GPI (34.5%) was significantly more frequent than aCL (13.6%) (p < 10⁻³). The frequency of aβ2GPI was significantly higher in patients with HT than in healthy population (34.5% vs 11.1%, p < 10⁻³). In HT patients, IgA isotype of aβ2GPI was significantly more common than in HBD and in GD patients (27.3% vs 7.8%, p < 10⁻³ and 27.3% vs 12.5%, p = 0.02, respectively).Conclusionaβ2GPI and not aCL were frequent in AITD. IgA was the predominant isotype of aβ2GPI. aβ2GPI‐IgA was more frequent in HT than in GD.     

Anticardiolipin and anti‐beta 2‐glycoprotein I antibodies in patients with unexplained articular manifestations

ObjectiveTo determine the frequency of antiphospholipid antibodies (aPL) in patients with unexplained articular manifestations.Material and MethodsThree hundred thirteen patients suffering from arthritis or arthralgia without evident cause and 266 healthy blood donors (HBD) were included in the study. Anticardiolipin antibodies (aCL) and anti‐beta 2‐glycoprotein I antibodies (aβ2GPI) were measured by ELISA.ResultOut of the 313 patients, 250 were females and 63 were males. The mean age of patients was 49 ± 14 years (17–87 years). One hundred eleven patients have arthralgia and 202 have arthritis. The frequency of aCL and/or aβ₂GPI (24.9%) was significantly higher in patients than in HBD (10.9%). The frequency of aβ2GPI was 23.6% in patients and 9.4% in the control group (p < 10⁻³). aβ2GPI‐IgA was significantly more frequent in patients than in the control group (20.4% vs. 7.5%, p < 10⁻³). aβ2GPI was most commonly observed than aCL in patients (23.6% vs. 6.4%, p < 10⁻⁶). IgA isotype of aβ2GPI was the most frequent in 20.4% of patients while IgG and IgM were detected in 5.4% and 2.9% respectively.ConclusionThis study showed that aPL were common in patients with articular manifestations and were mainly directed against β₂GPI. The role of these antibodies remains to be specified.     

Potential of long non‐coding RNA KCNQ1OT1 as a biomarker reflecting systemic inflammation,multiple organ dysfunction,and mortality risk in sepsis patients

BackgroundLong non‐coding RNA potassium voltage‐gated channel subfamily Q member 1 opposite strand 1 (lnc‐KCNQ1OT1) represses inflammation and multiple organ dysfunction, whereas its clinical value in sepsis is unclear. Thus, this study aimed to explore this issue.MethodsLnc‐KCNQ1OT1 from peripheral blood mononuclear cells were detected by RT‐qPCR in 116 sepsis patients and 60 healthy controls (HCs). Moreover, sepsis patients were followed‐up until death or up to 28 days.ResultsLnc‐KCNQ1OT1 decreased in patients with sepsis than in HCs (p < 0.001). In sepsis patients, lnc‐KCNQ1OT1 was negatively correlated with sequential organ failure assessment (SOFA) scores (r = −0.344, p < 0.001) and several SOFA subscale scores (including respiratory system, coagulation, liver, and renal systems) (all r < 0, p < 0.05). Furthermore, lnc‐KCNQ1OT1 was negatively correlated with CRP (r = −0.386, p < 0.001), TNF‐α (r = −0.332, p < 0.001), IL‐1β (r = −0.319, p < 0.001), and IL‐6 (r = −0.255, p = 0.006). Additionally, lnc‐KCNQ1OT1 levels were lower in sepsis deaths than in sepsis survivors (p < 0.001), and the receiver operating characteristic curve showed that lnc‐KCNQ1OT1 had an acceptable ability to predict 28‐day mortality (area under the curve: 0.780, 95% confidence interval: 0.678–0.882). Meanwhile, its ability to predict 28‐day mortality risk was higher than that of CRP, TNF‐α, IL‐1β, and IL‐6, but slightly lower than the SOFA score and acute physiology and chronic health evaluation II score.ConclusionLnc‐KCNQ1OT1 serves as a potential biomarker for monitoring disease severity and prognosis in patients with sepsis.     

Evaluating the potency of blood long noncoding RNA PVT1 as candidate biomarker reflecting inflammation,multiple organ dysfunction,and mortality risk in sepsis patients

ObjectiveLong noncoding RNA plasmacytoma variant translocation 1 (lnc‐PVT1) promotes septic inflammation and organ injuries via multiple ways, while its clinical engagement in sepsis management is indistinct. This study aimed to investigate its relationship with inflammation, multiple organ dysfunction, and mortality risk in sepsis patients.MethodsSepsis patients and age‐/gender‐matched healthy controls were enrolled; their lnc‐PVT1 expression in plasma were detected by RT‐qPCR. For sepsis patients only, the inflammatory cytokine levels (tumor necrosis factor (TNF)‐α, interleukin (IL)‐1β, and IL‐17A) in plasma were detected by ELISA. According to the survival data during 28‐day follow‐up, sepsis patients were divided into sepsis survivors and sepsis deaths.ResultsLnc‐PVT1 expression was increased in sepsis patients (N = 157) compared with healthy controls (N = 80) (p < 0.001). In sepsis patients, lnc‐PVT1 was linked with higher acute physiology and chronic health evaluation II (APACHEII) score (p = 0.001), total sequential organ failure assessment (SOFA) score, and its most subitems (SOFA‐respiratory system, SOFA‐coagulation, SOFA‐liver, SOFA‐cardiovascular system, and SOFA‐renal system scores) (all p < 0.01), but not SOFA‐nervous system score (p = 0.091); it did not relate to primary infection sites either (p = 0.204). Furthermore, lnc‐PVT1 correlated with increased C‐reactive protein, TNF‐α, IL‐1β, and IL‐17 in sepsis patients (all p < 0.01). Additionally, lnc‐PVT1 expression was higher in sepsis deaths than that in sepsis survivors (p < 0.001), following receiver‐operating characteristic curve disclosed that lnc‐PVT1 predicted 28‐day septic mortality risk (area under the curve: 0.789, 95% confidence interval: 0.702–0.875).ConclusionCirculating lnc‐PVT1 exhibits the potential as a biomarker in sepsis patients to inform inflammation, multiple organ dysfunction, and mortality risk.     

Transfusion‐induced platelet antibodies and regulatory T cells in multiply transfused patients

BackgroundPlatelet transfusion refractoriness (PTR) remains a difficult problem in patients requiring long‐term platelet supportive care. However, there are little data on the frequency of platelet antibodies in multiply transfused Chinese patients. Moreover, the relationship between peripheral regulatory T cells (Tregs) and PTR remains unclear.MethodsWe retrospectively studied the frequency of alloimmunization against platelet antigens in patients receiving multiple transfusions between 2013 and 2017. Monoclonal antibody solid‐phase platelet antibody test (MASPAT) kits were used to screen for platelet antibodies before each platelet transfusion. Peripheral Tregs and CD4⁺CD25⁺CD127⁻ T cells were detected by flow cytometry, while transforming growth factor‐beta (TGF‐β) and interleukin (IL)‐17 cytokines were detected by enzyme‐linked immunosorbent assay.ResultsA total of 399 patients who met the inclusion criteria were enrolled for the analysis of platelet antibodies and refractoriness. Among these patients, 10 (2.5%) were positive for platelet antibodies before transfusion and 47 (11.8%) became antibody‐positive during the study period. The number of alloimmunized patients was significantly higher in patients with hematological disease as compared with other disease groups (p < 0.05). Refractoriness and alloimmunization occurred in 77 (19.3%) and 22 (28.6%) patients, respectively. There were no significant differences in CD4⁺, CD8⁺, and CD4⁺CD25⁺CD127⁻ T cell numbers and plasma levels of TGF‐β1 and IL‐17 between patients with PTR and the control group.ConclusionsRefractoriness was common in patients undergoing multiple platelet transfusions (19.3%), with alloimmunization observed in 28.6% of patients. However, Tregs in peripheral blood may not play a key role in PTR.     

Association of tumor necrosis factor alpha ‐308 single nucleotide polymorphism with SARS CoV‐2 infection in an Iraqi Kurdish population

Uncovering risk factors playing roles in the severity of Coronavirus disease 2019 (Covid‐19) are important for understanding pathoimmunology of the disease caused by severe acute respiratory syndrome Coronavirus 2 (SARS CoV‐2). Genetic variations in innate immune genes have been found to be associated with Covid‐19 infections. A single‐nucleotide polymorphism (SNP) in a promoter region of tumor necrosis factor alpha (TNF‐α) gene, TNF‐α −308G>A, increases expression of TNF‐α protein against infectious diseases leading to immune dysregulations and organ damage. This study aims to discover associations between TNF‐α −308G>A SNP and Covid‐19 infection. Polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) was used for genotyping a general Kurdish population and Covid‐19 patients. The homozygous mutant (AA) genotype was found to be rare in the current studied population. Interestingly, the heterozygous (GA) genotype was significantly (p value = 0.0342) higher in the Covid‐19 patients than the general population. This suggests that TNF‐α −308G>A SNP might be associated with Covid‐19 infections. Further studies with larger sample sizes focusing on different ethnic populations are recommended.     

Elevated levels of interleukin‐35 and interleukin‐37 in adult patients with obstructive sleep apnea

BackgroundSystemic inflammation has a critical role in the pathogenesis of obstructive sleep apnea (OSA). Interleukin (IL)‐35 and IL‐37 have been identified as novel immune‐modulating cytokines with anti‐inflammatory activities in numerous types of inflammatory disease. The present study aimed to examine the serum levels of IL‐35 and IL‐37 in patients with OSA, and to investigate their associations with the severity of OSA.MethodsA total of 97 patients, including 67 cases of OSA and 30 age‐ and gender‐matched healthy control subjects, were enrolled in the present study. All subjects were evaluated by overnight polysomnography. Serum IL‐35, IL‐37, and pro‐inflammatory cytokine IL‐1β levels were examined by ELISA.ResultsCompared with those in the control subjects, serum IL‐35, IL‐37, and IL‐1β levels were significantly elevated in patients with mild, moderate, or severe OSA. Furthermore, a severity‐dependent increase in serum IL‐35 and IL‐37 levels was observed in patients with OSA. IL‐35 and IL‐37 levels were positively correlated with the apnea‐hypopnea index (r = 0.742 and 0.578, respectively; both p < 0.001), while they were negatively correlated with the mean oxygen saturation (r = −0.461 and −0.339, respectively; both p < 0.001) and lowest oxyhaemoglobin saturation (r = −0.616 and −0.463, respectively; both p < 0.001) in patients with OSA. In addition, a positive correlation was observed between IL‐35 or IL‐37 and IL‐1β levels (all p < 0.001).ConclusionThe serum levels of IL‐35 and IL‐37 were significantly increased in patients with OSA and associated with the severity of OSA, implying that IL‐35 and IL‐37 may have a protective role in OSA by counteracting inflammatory responses.     

CD8β Increases CD8 Coreceptor Function and Participation in TCR–Ligand Binding

To study the role of CD8β in T cell function, we derived a CD8α/β⁻ (CD8^−/−) T cell hybridoma of the H-2K^d–restricted N9 cytotoxic T lymphocyte clone specific for a photoreactive derivative of the Plasmodium berghei circumsporozoite peptide PbCS 252-260. This hybridoma was transfected either with CD8α alone or together with CD8β. All three hybridomas released interleukin 2 upon incubation with L cells expressing K^d–peptide derivative complexes, though CD8α/β cells did so more efficiently than CD8α/α and especially CD8^−/− cells. More strikingly, only CD8α/β cells were able to recognize a weak agonist peptide derivative variant. This recognition was abolished by Fab′ fragments of the anti-K^d α3 monoclonal antibody SF11.1.1 or substitution of K^d D-227 with K, both conditions known to impair CD8 coreceptor function. T cell receptor (TCR) photoaffinity labeling indicated that TCR–ligand binding on CD8α/β cells was ~5- and 20-fold more avid than on CD8α/a and CD8^−/− cells, respectively. SF1-1.1.1 Fab′ or K^d mutation D227K reduced the TCR photoaffinity labeling on CD8α/β cells to approximately the same low levels observed on CD8^−/− cells. These results indicate that CD8α/β is a more efficient coreceptor than CD8α/α, because it more avidly strengthens TCR–ligand binding.     

Identification of a Committed T Cell Precursor Population in Adult Human Peripheral Blood

Here, we report data concerning the discovery in adult human peripheral blood of a precursor cell population able to differentiate into CD4⁺CD3⁺αβ⁺ mature T cells. These cells, which represent 0.1–0.5% of total peripheral blood mononuclear cells (PBMC), express substantial levels of CD4, but lack CD3 surface expression. At a molecular level, they express the pre-T cell receptor α (pTα) gene, CD3-γ, CD-δ and CD-ε, and RAG-1 recombination enzyme and have initiated rearrangements in the T cell receptor (TCR)-β locus (D–J). Moreover, low levels of CD3ε protein, but not of TCR-β chain, can be detected in their cytoplasm. Our results suggest that CD4⁺CD3⁻ cells identified in peripheral blood are different from CD3⁻CD4⁺CD8⁻ thymocytes and may contain precursors of an extrathymic T cell differentiation pathway.     

Inter‐alpha‐trypsin inhibitor heavy chain 4: A serologic marker relating to disease risk,activity, and treatment outcomes of rheumatoid arthritis

ObjectiveInter‐alpha‐trypsin inhibitor heavy chain 4 (ITIH4) regulates immunity and inflammation, but its clinical role in rheumatoid arthritis (RA) patients remains unclear. Hence, this study was conducted to explore the association of circulating ITIH4 with disease risk, clinical features, inflammatory cytokines, and treatment outcomes of RA.MethodsAfter the enrollment of 93 active RA patients and 50 health controls (HCs), their serum ITIH4 level was analyzed by enzyme‐linked immunosorbent assay (ELISA). For RA patients only, serum ITIH4 level at week (W) 6 and W12 after treatment was also analyzed. Besides, serum tumor necrosis factor‐alpha (TNF‐α), interleukin (IL)‐1β, IL‐6, and IL‐17A at baseline of RA patients were also detected by ELISA.ResultsITIH4 was downregulated in RA patients (151.1 (interquartile range (IQR): 106.2–213.5) ng/mL) than in HCs (306.8 (IQR: 238.9–435.1) ng/mL) (p < 0.001). Furthermore, ITIH4 was negatively related to C‐reactive protein (CRP) (r_s  = −0.358, p < 0.001) and 28‐joint disease activity score using erythrocyte sedimentation rate (DAS28‐ESR) (r_s  = −0.253, p = 0.014) in RA patients, but not correlated with other clinical features (all p > 0.05). Besides, ITIH4 was negatively linked with TNF‐α (r_s  = −0.337, p = 0.001), IL‐6 (r_s  = −0.221, p = 0.033), and IL‐17A (r_s  = −0.368, p < 0.001) in RA patients, but not correlated with IL‐1β (r_s  = −0.195, p = 0.061). Moreover, ITIH4 was gradually elevated in RA patients from baseline to W12 after treatment (p < 0.001). Additionally, the increment of ITIH4 at W6 and W12 was linked with treatment response and remission in RA patients (all p < 0.05).ConclusionCirculating ITIH4 possesses clinical utility in monitoring disease risk, inflammation, disease activity, and treatment outcomes of RA.