Objective To prepare mouse monoclonal antibodies against the ectodomain of E2 (E2ecto) glycoprotein of Western equine encephalitis virus (WEEV). Methods A prokaryotic expression plasmid pET-28a-WEEV E2ecto was constructed and transformed into BL21 (DE3) competent cells. E2ecto protein was expressed by IPTG induction and presented mainly as inclusion bodies. Then the purified E2ecto protein was prepared by denaturation, renaturation and ultrafiltration. BALB/c mice were immunized with the formulated E2ecto protein using QuickAntibody-Mouse5W as an adjuvant via intramuscular route, boosted once at an interval of 21 days. At 35 days post-immunization, mice with antibody titer above 1×10⁴ were inoculated with E2ecto intraperitoneally, and spleen cells were fused with SP2/0 cells three days later. Hybridoma cells secreting specific monoclonal antibodies were screened by the limited dilution method, and ascites were prepared after intraperitoneal inoculation of hybridoma cells. The subtypes and titers of the antibodies in ascites were assayed by ELISA. The biological activity of the mAb was identified by immunofluorescence assay(IFA) on BHK-21 cells which were transfected with eukaryotic expression plasmid pCAGGS-WEEV-CE3E2E1. The specificity of the antibodies were evaluated with E2ecto proteins from EEEV and VEEV. Results Purified WEEV E2ecto protein was successfully expressed and obtained. Four monoclonal antibodies, 3G6G10, 3D7G2, 3B9E8 and 3D5B7, were prepared, and their subtypes were IgG2c(κ), IgM(κ), IgM(κ) and IgG1(κ), respectively. The titers of ascites antibodies 3G6G10, 3B9E8 and 3D7G2 were 10⁵, and 3D5B7 reached 10⁷. None of the four antibody strains cross-reacted with other encephalitis alphavirus such as VEEV and EEEV. Conclusion Four strains of mouse mAb specifically binding WEEV E2ecto are successfully prepared.
Horses ; Animals ; Mice ; Encephalitis Virus, Western Equine ; Ascites ; Immunosuppressive Agents ; Antibodies, Monoclonal ; Immunoglobulin M

To investigate the risk factors of poor prognosis and recurrence in patients with anti-NMDAR encephalitis. A single center, observational cohort study was used to retrospectively analyze 44 patients with anti NMDAR encephalitis hospitalized in the Department of Neurology of Beijing Tong Ren Hospital from January 2014 to October 2020. The results showed that the interval from onset to immunotherapy in the poor prognosis group was significantly longer than that in the good prognosis group (t=2.045,P=0.047), and the course of disease in the poor prognosis group was significantly longer than that in the good prognosis group (t=4.127,P=0.000 2). The number of patients with clinical manifestations of dyskinesia was significantly increased (Fisher exact test: P=0.014). The patients with abnormal brain MRI in the poor prognosis group were significantly more than those in the good prognosis group (Fisher exact test: P=0.017), and the patients with slow wave>50% in the poor prognosis group were significantly more than those with slow wave <50% (Fisher exact test: P<0.001). Patients with the first onset of immunotherapy time <3 months, long course of disease, high intracranial pressure, and high cerebrospinal fluid protein are prone to relapse. Bivariate logistic regression analysis showed that patients with dyskinesia, abnormal brain MRI, and slow wave EEG more than 50% were risk factors for poor prognosis (OR values were 4.687, 4.978, and 24.500, respectively; P values were 0.018, 0.016, and 0.000, respectively). The time of first-line immunotherapy for the first onset<3 months was the risk factor for recurrence (OR 17.231, P=0.010). In conclusion, dyskinesia, abnormal brain MRI and slow wave of EEG more than 50% may be the risk factors for poor prognosis of patients. The duration of immunotherapy less than 3 months after the first onset might be the risk factor for recurrence.
Humans ; Anti-N-Methyl-D-Aspartate Receptor Encephalitis/cerebrospinal fluid* ; Retrospective Studies ; Neoplasm Recurrence, Local ; Risk Factors ; Dyskinesias

OBJECTIVES: To study the clinical features of children with autoimmune encephalitis (AE) secondary to epidemic encephalitis B (EEB). METHODS: A retrospective analysis was performed on the medical data of five children with EEB with "bipolar course" who were treated in Children's Hospital Affiliated to Zhengzhou University from January 2020 to June 2022. RESULTS: Among the five children, there were three boys and two girls, with a median age of onset of 7 years (range 3 years 9 months to 12 years) and a median time of 32 (range 25-37) days from the onset of EEB to the appearance of AE symptoms. The main symptoms in the AE stage included dyskinesia (5/5), low-grade fever (4/5), mental and behavioral disorders (4/5), convulsion (2/5), severe disturbance of consciousness (2/5), and limb weakness (1/5). Compared with the results of cranial MRI in the acute phase of EEB, the lesions were enlarged in 3 children and unchanged in 2 children showed on cranial MRI in the AE stage. In the AE stage, four children were positive for anti-N-methyl-D-aspartate receptor antibody (one was also positive for anti-γ-aminobutyric acid type B receptor antibody), and one was negative for all AE antibodies. All five children in the AE stage responded to immunotherapy and were followed up for 3 months, among whom one almost recovered and four still had neurological dysfunction. CONCLUSIONS EEB can induce AE, with anti-N-methyl-D-aspartate receptor encephalitis as the most common disease. The symptoms in the AE stage are similar to those of classical anti-N-methyl-D-aspartate receptor encephalitis. Immunotherapy is effective for children with AE secondary to EEB, and the prognosis might be related to neurological dysfunction in the acute phase of EEB.
Male ; Female ; Humans ; Child ; Infant, Newborn ; Anti-N-Methyl-D-Aspartate Receptor Encephalitis ; Retrospective Studies ; Hashimoto Disease/therapy* ; Encephalitis, Arbovirus

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is an autoimmune inflammatory disease of the central nervous system, and little is known about its immune mechanism at present. There is a lack of disease-related biomarkers in cerebrospinal fluid except anti-NMDAR antibody, which leads to delayed diagnosis and treatment in some patients. Therefore, there has been an increasing number of studies on related cytokines in recent years to assess whether they can be used as new biomarkers for evaluating disease conditions and assisting diagnosis and treatment. Current studies have shown that some cytokines may be associated with the progression of anti-NMDAR encephalitis, and this article reviews the research advances in such cytokines associated with anti-NMDAR encephalitis.
Humans ; Cytokines ; Anti-N-Methyl-D-Aspartate Receptor Encephalitis/therapy* ; Biomarkers

Child ; Humans ; Encephalitis/therapy* ; Hashimoto Disease/therapy* ; Autoimmune Diseases of the Nervous System/diagnosis*

Objective: To evaluate the value of the 2020 diagnostic criteria (Cellucci criteria) for pediatric autoimmune encephalitis (AE) in children with suspected AE in a single center. Methods: The clinical data of 121 children hospitalized at the First Affiliated Hospital of Zhengzhou University from October 2019 to October 2021, with a diagnosis of suspected AE, were retrospectively collected and analyzed. The children were divided into definite antibody-positive AE (dAPAE), probable antibody-negative AE (prANAE), possible AE (pAE) and non-AE groups according to the Chinese expert consensus and the Graus criteria. A new diagnosis was made according to the Cellucci criteria which was compared with the clinical diagnosis to evaluate the diagnostic value of the Cellucci criteria. The Mann-Whitney U test, Kruskal-Wallis test, and χ² test were used to compare the differences among groups. The sensitivity and specificity were used to evaluate efficacy of the Cellucci criteria. Results: Among the 121 children, 72 were males and 49 were females, with an age of 10.3 (6.5, 14.0) years at disease onset. There were 99 cases diagnosed as AE according the clinical diagnosis (58 males and 41 females), of which 43 cases were diagnosed as dAPAE, 14 cases as prANAE and 42 cases as pAE, and the other 22 cases were not AE (14 males and 8 females). The top 2 initial symptoms in the 99 children with AE were seizures (53 cases, 53.5%) and abnormal mental behaviors (35 cases, 35.4%). And the most common symptoms during the course of the disease were abnormal mental behaviors (77 cases, 77.8%) and seizures (64 cases, 64.6%). There were statistically differences in the incidence of consciousness disorders, autonomic dysfunctions during the course of the disease and the length of hospitalization among the 4 groups (χ²=21.63, 13.74, H=22.60, all P<0.05). Ninety-six of the 121 children were tested for AE-related antibodies, of which 45 cases (46.9%) were antibody-positive. According to the Cellucci criteria, 42 cases were diagnosed as dAPAE, 34 cases as prANAE and 14 cases as pAE. Compared with the clinical diagnosis, the sensitivity of the Cellucci criteria for the diagnosis of the 3 types of AE were 93.02%, 92.86% and 87.88%, and the specificity were 96.23%, 74.39% and 86.36%, respectively. Conclusions: The Cellucci criteria has a high sensitivity and specificity for the diagnosis of pAE and dAPAE in the clinical management of children with suspected AE, while a high sensitivity but low specificity for the diagnosis of prANAE. Therefore, it is recommended to apply the Cellucci criteria selectively in clinical practice according to the actual situation, especially in the diagnosis of prANAE.
Child ; Encephalitis/diagnosis* ; Female ; Hashimoto Disease/diagnosis* ; Humans ; Male ; Retrospective Studies ; Seizures

Aminobutyrates ; Autoantibodies ; Child ; Encephalitis ; Humans

Humans ; Encephalitis

Objective: To summarize the clinical characteristics, diagnosis and treatment of forest encephalitis, and provide basis for revising relevant diagnostic criteria. Methods: From January to December 2020, the clinical characteristics, diagnosis and treatment of forest encephalitis cases in the data of China National Knowledge Infrastructure (CNKI) and Wanfang Chinese journals from 2009 to 2020 were retrospectively analyzed. The measurement data are expressed in Mean±SD, numbers, and the counting data of gender, region and occupation are expressed in numbers and composition ratio. Descriptive analysis of relevant data is carried out. Results: There were 1 153 confirmed cases of forest encephalitis reported in domestic literature, including 910 males and 243 females. Age: 16-78 years old; Cases were mainly distributed in Jilin Province, Inner Mongolia Autonomous Region and Heilongjiang Province. The cases included forest rangers (112/518), freelancers (104/518) and loggers (88/518). The common symptoms and signs were fever 81.2% (936/1153), headache 70.3% (811/1153), meninges irritation 29.0% (334/1153), vomiting 25.3% (292/1153), nausea 21.8% (251/1153), etc. Only 48.1% (555/1153) of the patients obtained positive pathogenic test results from blood and/or cerebrospinal fluid specimens. 42.1% (485/1153) had definite diagnosis grade, 354 cases were mild, 58 cases were moderate, and 73 cases were severe. Among 730 patients with forest encephalitis who received complete treatment, 511 cases were cured, 148 cases were improved, 48 cases were not cured, and 23 cases died. Conclusion: The epidemic of forest encephalitis has strict regional, seasonal and occupational characteristics. Early diagnosis and treatment can reduce the mortality and disability rate.
Male ; Female ; Humans ; Adolescent ; Young Adult ; Adult ; Middle Aged ; Aged ; Retrospective Studies ; Encephalitis, Tick-Borne ; Encephalitis/epidemiology* ; Headache ; Fever

Introduction: The etiology of encephalitis involves an enormous range and can be classified as infectious or immune-mediated. There are several factors influencing its prognosis and has been associated with significant morbidity and mortality. This study aims to evaluate the clinico-epidemiologic characteristics and outcomes of infectious and immune-mediated encephalitis among pediatric patients. Methodology: Retrospective descriptive cross-sectional study that included patients aged 6 months to 17 years old with encephalitis in a tertiary hospital between January 2010 to December 2020 Results: A total of 23 cases were reviewed and 60.87% were infectious while that of immune-mediated was 39.13%. Among those with identified infectious cause, Mycoplasma pneumonia was the most common (28.57%). Infectious encephalitis was more common among younger males (35.71%) while immune-mediated affected female adolescents more (55.56%). The most common neurologic manifestation was altered mental status and/or behavioral changes. Treatment such as antibiotics (78.26%), anticonvulsant therapy (78.26%), and steroids (43.48%) were given. All immune-mediated cases received steroids. More than half of patients had complete recovery (56.52%). Conclusion Pediatric encephalitis should be considered among patients with neurologic dysfunction with or without systemic involvement. Behavioral changes in an apparently well child should prompt clinicians to consider anti-NMDAR encephalitis, especially if viral studies are negative and with no other known cause. Viruses remain to be the most common etiology, but other possible causes should be highly considered such as anti-NMDAR and Mycoplasma. A normal CSF analysis, imaging and/or encephalography (EEG) may not totally exclude encephalitis. Prognosis is relatively good hence an early diagnosis and initiation of appropriate management is important.
Encephalitis ; Infectious Encephalitis ; Encephalitis, Viral ; Mycoplasma