OBJECTIVE: To analyze the Epstein-Barr virus (EBV) load in different lymphocyte subsets, as well as clinical characteristics and outcomes in patients with hematologic malignancies experiencing EBV reactivation. METHODS: Peripheral blood samples from patients were collected. B, T, and NK cells were isolated sorting with magnetic beads by flow cytometry. The EBV load in each subset was quantitated by real-time quantitative polymerase chain reaction (RT-qPCR). Clinical data were colleted from electronic medical records. Survival status was followed up through outpatient visits and telephone calls. Statistical analyses were performed using SPSS 25.0. RESULTS: A total of 39 patients with hematologic malignancies were included, among whom 35 patients had undergone allogeneic hematopoietic stem cell transplantation (allo-HSCT). The median time to EBV reactivation was 4.8 months (range: 1.7-57.1 months) after allo-HSCT. EBV was detected in B, T, and NK cells in 20 patients, in B and T cells in 11 patients, and only in B cells in 4 patients. In the 35 patients, the median EBV load in B cells was 2.19×10⁴ copies/ml, significantly higher than that in T cells (4.00×10³ copies/ml, P <0.01) and NK cells (2.85×10² copies/ml, P <0.01). Rituximab (RTX) was administered for 32 patients, resulting in EBV negativity in 32 patients with a median time of 8 days (range: 2-39 days). Post-treatment analysis of 13 patients showed EBV were all negative in B, T, and NK cells. In the four non-transplant patients, the median time to EBV reactivation was 35 days (range: 1-328 days) after diagnosis of the primary disease. EBV was detected in one or two subsets of B, T, or NK cells, but not simultaneously in all three subsets. These patients received a combination chemotherapy targeting at the primary disease, with 3 patients achieving EBV negativity, and the median time to be negative was 40 days (range: 13-75 days). CONCLUSION In hematologic malignancy patients after allo-HSCT, EBV reactivation commonly involves B, T, and NK cells, with a significantly higher viral load in B cells compared to T and NK cells. Rituximab is effective for EBV clearance. In non-transplant patients, EBV reactivation is restricted to one or two lymphocyte subsets, and clearance is slower, highlighting the need for prompt anti-tumor therapy.
Humans ; Hematologic Neoplasms/virology* ; Herpesvirus 4, Human/physiology* ; Epstein-Barr Virus Infections ; Hematopoietic Stem Cell Transplantation ; Virus Activation ; Lymphocyte Subsets/virology* ; Flow Cytometry ; Killer Cells, Natural/virology* ; Male ; Female ; B-Lymphocytes/virology* ; Viral Load ; Adult ; T-Lymphocytes/virology* ; Middle Aged

OBJECTIVETo study the clinical and laboratory characteristics of chronic active Epstein-Barr virus (EBV) infection (CAEBV) in children and to provide a basis for the diagnosis and treatment of CAEBV.

METHODSThe clinical data of 13 children with CAEBV, as well as 15 cases of acute EBV infection (AEBV) as controls, were analyzed, including clinical manifestations, EBV antibodies, EBV DNA, and peripheral blood lymphocyte subsets.

RESULTSBoth groups of patients had infectious mononucleosis-like symptoms such as fever, hepatomegaly, splenomegaly, and lymphadenectasis, but CAEBV patients had a longer course of disease and continuous and recurrent symptoms. Compared with the AEBV group, the CAEBV group had a significantly higher EBV DNA load in peripheral blood (P<0.05), a significantly higher VCA-IgG titer (P<0.05), and significantly lower numbers of white blood cells, lymphocytes, B cells, total T cells, CD4+ T cells, and CD8+ T cells in peripheral blood (P<0.05). Among 13 CAEBV patients followed up, 8 cases died, 2 cases showed an improvement, 2 cases had a recurrence, and 1 case was lost to follow-up after being transferred to another hospital. All the AEBV patients were cured and had no recurrence during the one-year follow-up.

CONCLUSIONSThe clinical manifestations of CAEBV vary in children. It is difficult to distinguish CAEBV from AEBV early. More attention should be paid to CAEBV because of its severe complications, poor prognosis, and high mortality. Measurement of EBV DNA load, VCA-IgG titer, and lymphocyte subsets in peripheral blood may be helpful in the diagnosis and differential diagnosis of CAEBV.

Adolescent ; Child ; Child, Preschool ; Chronic Disease ; Epstein-Barr Virus Infections ; diagnosis ; immunology ; virology ; Female ; Humans ; Infant ; Lymphocyte Subsets ; immunology ; Male

The protective efficacy of DNA plasmids encoding avian infectious bronchitis virus (IBV) S1, N, or M protein was investigated in chickens. Chickens were inoculated monovalently (with plasmid pVAX1-16S1, pVAX1-16M, or pVAX1-16N alone) or multivalently (combination of the three different plasmids, pVAX1-16S1/M/N). A prime-boost immunization protocol against IBV was developed. Chickens were immunized with the multivalent DNA vaccine twice and then boosted with an inactivated vaccine once. Antibody titers of the chickens immunized with pVAX1-16S1/M/N were much higher than those of the monovalent groups (p < 0.01). A protective rate up to 90% was observed in the pVAX1-16S1/M/N group. The serum antibody titers in the prime-boost birds were significantly higher than those of the multivalent DNA vaccine group (p < 0.01) but not significantly different compared to the inactivated vaccine group at 49 days of age. Additionally, the prime-boost group also showed the highest level of IBV-specific cellular proliferation compared to the monovalent groups (p < 0.01) but no significant difference was found compared to the multivalent DNA vaccine group, and the prime-boost group completely protected from followed viral challenge.
Aging ; Animals ; Antibodies, Viral/blood ; Cell Proliferation ; Chickens ; Coronavirus Infections/prevention & control/*veterinary/virology ; Immunization, Secondary/veterinary ; Infectious bronchitis virus/*immunology ; Poultry Diseases/*prevention & control/virology ; T-Lymphocyte Subsets/cytology/physiology ; Vaccines, DNA/immunology ; Vaccines, Inactivated/immunology ; Viral Vaccines/*immunology

OBJECTIVESTo study the effect of HAART on subsets of T lymphocytes and expression of CD127 on memory and naїve CD4(+) and CD8(+)T cells in pediatric AIDS patients with different viral loads receiving HAART.

METHODA cross- sectional study on 194 pediatric AIDS patients receiving HAART was carried out and 52 age matched healthy children were recruited as controls. The percentage of CD4(+), CD8(+), CD8(+)CD45RA(+)CD127(+/-), CD8(+)CD45RO(+)CD127(+/-), CD4(+)CD45RA(+)CD127(+/-) and CD4(+)CD45RO(+)CD127(+/-)T cells was tested using flow cytometry, and HIV-RNA in plasma was detected by quantitative RT-PCR.

RESULTThe percentage of memory (CD45RO(+)) CD4(+)T cells decreased to (45.73 ± 8.85)%, and that of naїve (CD45RA(+)) CD4(+) and memory CD8(+)T increased to (60.44 ± 5.01)% and (54.69 ± 7.71) % respectively in the pediatric AIDS patients vs. controls (P < 0.05). The percentage of naїve (CD45RA(+)) CD4(+)T cells of patients with viral load (VL) < 400 copies/ml was (65.57 ± 5.33) %, which was significantly higher than that of patients with VL ≥ 400 copies/ml (P < 0.05).Of patients with VL < 400 copies/ml, the percentage of CD4(+)CD127(+)T cells, especially the subset of memory CD4(+)CD127(+)T cells was (82.35 ± 2.31)%, which was higher than that of patients with VL ≥ 400 copies/ml, but lower than that of controls (P < 0.05). The percentage of memory and naїve CD8(+)CD127(+)T cells was lower than that of controls (P < 0.05).

CONCLUSIONThe recovery of CD4(+)T cell subsets in pediatric AIDS patients is associated with viral load. Effective HAART can increase the percentage of naїve CD4(+)T cells and the life of memory CD4(+)T cells.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; virology ; Adolescent ; Antiretroviral Therapy, Highly Active ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Cross-Sectional Studies ; Female ; Flow Cytometry ; Humans ; Immunologic Memory ; Interleukin-7 Receptor alpha Subunit ; immunology ; metabolism ; Lymphocyte Count ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocyte Subsets ; immunology ; Viral Load

OBJECTIVETo investigate the effect of human cytomegalovirus (HCMV) infection on T lymphocyte subsets in children with β-thalassemia major (TM) during the initial 6 months after allogeneic hematopoietic stem cell transplantation (Allo-HSCT).

METHODSFrom January, 2010 to January, 2011, 35 children with TM underwent Allo-HSCT. Peripheral blood samples were obtained from the children 6 month after the transplantation to examine the changes of T lymphocytes subsets in relation to HCMV seropositivity.

RESULTSThirteen children were found seropositive and 22 were seronegative for HCMV. The HCMV-seropositive children had a higher CD8⁺ cell percentage but a lower CD4⁺ cell percentage than those without HCMV infection. Compared with those seronegative for HCMV, the children with HCMV seropositivity showed increased percentages of CD8⁺ cells and CD8⁺CD28⁻ cells with a decreased percentage of CD8⁺CD28⁺ cells. A positive linear correlation was found between the percentages of CD8⁺CD28⁻ cells and CD8⁺ cells.

CONCLUSIONHCMV infection can lead to the accumulation of CD8⁺CD28 cells to cause increased CD8⁺ T cells in the peripheral blood in TM children after Allo-HSCT. The percentages of CD8⁺CD28⁻ cells has a positive linear correlation to that of CD8⁺ cells.

Adolescent ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Child, Preschool ; Cytomegalovirus ; Cytomegalovirus Infections ; immunology ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Postoperative Period ; T-Lymphocyte Subsets ; beta-Thalassemia ; immunology ; surgery ; virology

Natural killer (NK) cells play an important role in innate immunity, especially in the response to viral infections, such as hepatitis C virus (HCV). Killer cell immunoglobulin-like receptors (KIRs) are the primary receptors of NK cells that mediate innate immunity. KIRs are also involved in acquired immunity, because some KIRs are expressed on the surface of certain subsets of T cells. In this study, the frequency of KIR genes, HLA-C allotypes, and combinations of KIR genes with their HLA-C ligands were evaluated in two different groups of the Korean population: controls and patients with chronic HCV infection. The study population consisted of 147 Korean patients with chronic HCV infection. The frequency of KIR2DS2 in patients with chronic HCV infection was 9.5% which was significantly lower than 19.5% of the control (P < 0.01). However, there were no significant differences in the frequency of other KIR genes, HLA-C allotypes or different combinations of KIR genes with their HLA-C ligands. This study can contribute to the further prospective study with a larger scale, suggesting the assumption that KIR2DS2 might aid in HCV clearance by enhancing both the innate and acquired immune responses of people in Korea.
Adult ; Aged ; Female ; Genes, MHC Class I ; Genotype ; HLA-C Antigens/genetics ; Hepacivirus/immunology ; Hepatitis C, Chronic/*genetics/immunology ; Humans ; Killer Cells, Natural/immunology/virology ; Male ; Middle Aged ; Receptors, KIR/*genetics/immunology ; Republic of Korea ; T-Lymphocyte Subsets/immunology

OBJECTIVETo observe the effect of Shugan Jianpi Bushen Recipe (SJBR) on the splenic T lymphocytes and virus load in the hepatitis B virus (HBV) transgenic (Tg) mice, and to study its antiviral efficacy and mechanisms of action.

METHODSSixty male BALB/C mice of SPF grade were included. Ten non-HBV Tg male mice were included as the normal control group. Fifty HBV Tg mice were randomly divided into five groups, i. e., the model group, the adefovir (ADV) group, the low dose SJBR group, the middle dose SJBR group, and the high dose SJBR group, ten in each. 10, 20, and 40 g/kg SJBR crude drug was respectively given by gastro-gavage to mice in the low dose SJBR group, the middle dose SJBR group, and the high dose SJBR group. ADV 50 mg/kg body weight was given by gastrogavage to mice in the ADV group. Equal volume of sterilized iso-osmia was given to mice in the normal group and the model group. The medication was performed once daily, totally for 21 successive days. The serum HBV DNA titers of HBV Tg mice were detected using Real-time fluorescent PCR one day before administration (T0), ten days after administration (T1), 21 days after administration (T2), and three days after withdrawal (T3), respectively; the serum hepatitis B surface antigen (HBsAg) of HBV Tg mice on T3 was detected by ELISA. The splenic T lymphocyte percent of all mice was detected by flow cytometry.

RESULTSSerum HBsAg at TO was positive in the high-, middle-, low-dose SJBR, and ADV groups. The HBsAg negative rate at T3 was lower in the high dose SJBR group than in the ADV group, showing statistical difference (P<0.01). Compared with TO of the same group, the serum HBV DNA titers could be continually decreased by high dose SJBR, showing statistical difference (P<0.01). The serum HBV DNA titers also gradually decreased in the ADV group (P<0.01), but it somewhat increased at T3. The CD3+ cell percent could be elevated by high-, middle-, low-dose SJBR, and ADV groups (P<0.05, P<0.01). The CD8+ T cell percent could also be obviously lowered by high-, middle-, and low-dose SJBR (P<0.01). Compared with the middle-, low-dose SJBR, and ADV groups, the CD4+ T cell percent and CD4+/CD8+ increased as well as CD8+ decreased in the high dose SJBR group, showing statistical difference (P<0.01). The CD3+ T cell percent was significantly positively correlated to the decrement of HBV DNA titers between the pre-treatment and post-treatment in the middle dose SJBR group (r=0.654, P<0.05). The percents of CD4+, CD8+ T cells and CD4+/CD8+ were significantly positively correlated to the decrement of HBV DNA titers between the pre-treatment and post-treatment in the high dose SJBR group (r=0.53, r=0.79, r =0.80, P<0.01).

CONCLUSIONSSJBR were capable of inhibiting the HBV DNA duplication of HBV Tg mice. One of its anti-HBV mechanisms possibly be improving the the abnormality of T lymphocyte subsets and the immune function.

Adenine ; analogs & derivatives ; pharmacology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hepatitis B virus ; drug effects ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Organophosphonates ; pharmacology ; Spleen ; cytology ; virology ; T-Lymphocyte Subsets ; drug effects ; Viral Load ; drug effects ; Virus Replication ; drug effects

OBJECTIVETo investigate the relationship between coxsackievirus infection and type 1 diabetes mellitus (T1DM), and observe the changes of T lymphocyte subsets in the development of T1DM.

METHODSWe detected Coxsackievirus RNA by reverse transcription PCR, and measured the change in T-lymphocyte subsets by flow cytometry in 22 cases of newly diagnosed T1DM (group I), 30 patients with diabetes for some time (group II), and 30 healthy subjects (group III).

RESULTSThe positivity rate of coxsackie virus RNA in groups I, II, and III was 55.55%, 23.33%, and 6.67%, respectively, showing a significant difference among the 3 groups (P<0.01). Patients with upper respiratory tract infection had a higher positivity rate for coxsackie virus RNA than those without upper respiratory tract infection in group I (P<0.05). Compared with the control group, the percentage of CD3, CD4 and CD4/CD8 ratio decreased significantly in groups I and II (P<0.01 or P<0.05). CD3, CD4 and CD4/CD8 tended to increase in group II in comparison with group I, and there was an significant difference in CD3 and CD4 between the two groups (P<0.01 or P<0.05). Compared with the control group and CVBRNA-negative group, CVBRNA-positive group showed significantly lowered CD3, CD4, CD8 and CD4/CD8 (P<0.01 or P<0.05).

CONCLUSIONThe occurrence and development of type 1 diabetes is closely related to coxsackie virus infection, and the changes in T lymphocyte subsets serves as a probable mechanism of its pathogenicity.

Adolescent ; Adult ; CD4-CD8 Ratio ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Coxsackievirus Infections ; complications ; immunology ; Diabetes Mellitus, Type 1 ; complications ; virology ; Female ; Humans ; Lymphocyte Count ; Male ; T-Lymphocyte Subsets ; immunology ; Young Adult

Administration, Oral ; Anti-Bacterial Agents ; administration & dosage ; Cytomegalovirus Infections ; drug therapy ; immunology ; virology ; Erythromycin ; administration & dosage ; Female ; Humans ; Infant ; Male ; T-Lymphocyte Subsets ; immunology ; Virus Replication

OBJECTIVETo investigate role of CD4-CD8- T cells in murine hepatitis virus type 3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice and to identify their surface markers.

METHODSThirty C3H/Hej mice received 10 Pfu MHV-3 intraperitoneally, the CD4-CD8- T cells were isolated using magnetic bead sorting on 0, 4, 15, 30, 40 days post MHV-3 infection. The cytotoxic effects of CD4-CD8- T cells on normal and infected hepatocytes, CD8+ T cells and unrelated-virus (murine cytomegalovirus, MCMV) infected CD8+ T cells were examined by non-radioactive cytotoxicity assay. The surface markers of CD4-CD8- T cells were determined by flow cytometry.

RESULTSMHV-3 infected CD4-CD8- T cells showed significant cytotoxic effect on CD8+ T cells, but not on infected hepatocytes or MCMV infected CD8+ T cells. The analysis of cell surface markers demonstrated that the CD4-CD8- T cells are a completely new T cell subset.

CONCLUSIONSCD4-CD8- T cells have significant cytotoxic effect on virus specific CD8+ T cells in MHV-3 infected C3H/Hej mice, which suggests that CD4-CD8- T cells have immune modulatory functions in the development of chronic viral hepatitis. The phenotype of these CD4-CD8- T cells detected by flow cytometry is TCR alpha beta +CD3+CD4- CD8- CD25- CD28- CD30- CD44+.

Animals ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Coronavirus Infections ; immunology ; pathology ; virology ; Female ; Flow Cytometry ; Hepatitis, Viral, Animal ; immunology ; pathology ; virology ; Liver ; immunology ; pathology ; Mice ; Mice, Inbred C3H ; Murine hepatitis virus ; Spleen ; immunology ; pathology ; T-Lymphocyte Subsets ; immunology ; Time Factors