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1.
Journal of Experimental Hematology ; (6): 1429-1435, 2021.
Article in Chinese | WPRIM | ID: wpr-922276

ABSTRACT

OBJECTIVE@#To establish the in vivo traceable acute myeloid leukemia mice model with Luciferase-Expressing KG1a Cells.@*METHODS@#KG1a cells with stable luciferase gene expression (called as KG1a-Luc cells) were constructed by lentivirus transfection, then sifted out by puromycin. Eighteen male NOD-SCID-IL2rg@*RESULTS@#KG1a cells expressing luciferase stably were successfully obtained. The tumor luminescence wildly spread at day 17 captured by in vivo imaging. The KG1a-Luc tumor cells could be detected in the peripheral blood of the mice, with the average percentage of (16.27±6.66)%. The morphology and pathology result showed that KG1a-Luc cells infiltrate was detected in bone marrow, spleens and livers. The survival time of the KG1a-Luc mice was notably shorter as compared with those in the control group, the median survival time was 30.5 days (95%CI: 0.008-0.260).@*CONCLUSION@#The acute myeloid leukemia NOD-SCID-IL2rg


Subject(s)
Animals , Disease Models, Animal , Interleukin Receptor Common gamma Subunit , Leukemia, Myeloid, Acute , Luciferases/genetics , Male , Mice , Mice, Inbred NOD , Mice, SCID
2.
Article in Chinese | WPRIM | ID: wpr-772338

ABSTRACT

BACKGROUND@#Small cell lung cancer (SCLC) is characterized by poor differentiation, high malignancy and rapid growth fast, short double time, early and extensive metastatic malignancy. In clinical, chemotherapy is the main treatment method, while resistance to multiple chemotherapy drugs in six to nine months has been a major clinical challenge in SCLC treatment. Therefore, It has important clinical value to building SCLC aninimal model which is similar to patients with SCLC. Animal model of xenotransplantation (PDX) from the patients with small cell lung cancer can well retain the characteristics of primary tumor and is an ideal preclinical animal model. The study is aimed to establish SCLC PDX animal model and induce the chemoresistance model to help to study the mechanism of chemoresistance and individual treatment.@*METHODS@#Fresh surgical excision or puncture specimens from SCLC patients were transplanted into B-NSGTM mice subcutaneous tissues with severe immunodeficiency in one hour after operation the B-NSGTM mice subcutaneous in 1 hour, and inject chemotherapy drugs intraperitoneally after its tumor growed to 400 mm³ with EP which is cisplatin 8 mg/kg eight days and etoposide 5 mg/kg every two days until 8 cycles. Measure the tumor volum and mice weights regularly, then re-engrafted the largest tumor and continue chemotherapy.@*RESULTS@#Nine cases were conducted for B-NSG mice modeling. Three of nine cases could be engrafted to new B-NSG mice at least two generation. The SCLC PDX animal models have been established successfully. After adopting chemotherapy drugs, the chemoresistance PDX models have been established. High homogeneity was found between xenograft tumor and patient's tumor in histopathology, immunohistochemical phenotype (Syn, CD56, Ki67).@*CONCLUSIONS@#The SCLC PDX animal model and the chemoresistance PDX animal model have been successfully constructed, the success rate is 33%, which provides a platform for the clinical research, seeking for biological markers and choosing individual treatment methods of SCLC.


Subject(s)
Animals , Antineoplastic Combined Chemotherapy Protocols , Pharmacology , Cisplatin , Disease Models, Animal , Drug Resistance, Neoplasm , Etoposide , Female , Humans , Interleukin Receptor Common gamma Subunit , Genetics , Lung Neoplasms , Drug Therapy , Metabolism , Pathology , Mice, Inbred BALB C , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Small Cell Lung Carcinoma , Drug Therapy , Metabolism , Pathology , Transplantation, Heterologous , Methods , Xenograft Model Antitumor Assays
3.
Article in Chinese | WPRIM | ID: wpr-689640

ABSTRACT

This research investigated the clinical features of immunodeficiency disease and the features of the mutation of its pathogenic genes. All 7 patients were boys aged 5 months to 4 years and 6 months and had a history of recurrent respiratory infection and pneumonia, low levels of IgM and IgG, and abnormal absolute values or percentages of lymphocyte subsets. High-throughput sequencing showed c.1684C>T mutations in the BTK gene in patient 1 and IVS8+2T>C splice site mutations in the BTK gene in patient 2. Both of these mutations came from their mothers. Patients 3, 4, and 5 had mutations in the IL2RG gene, i.e., c.298C>T, IVS3-2A>G, and c.164T>A, among which c.164T>A mutations had not been reported. Patient 6 had c.204C>G mutations in the RAG2 gene. Patient 7 had complex heterozygous mutations of c.913C>T and c.824G>A in the RAG2 gene, which came from his father and mother, respectively. Patients with immunodeficiency disease have abnormal immunological indices, and high-throughput sequencing helps to make a definite diagnosis.


Subject(s)
Agammaglobulinaemia Tyrosine Kinase , Agammaglobulinemia , Genetics , Child, Preschool , Computational Biology , DNA-Binding Proteins , Genetics , Genetic Diseases, X-Linked , Genetics , High-Throughput Nucleotide Sequencing , Humans , Immunologic Deficiency Syndromes , Genetics , Therapeutics , Infant , Interleukin Receptor Common gamma Subunit , Genetics , Male , Mutation , Nuclear Proteins , Genetics , Protein-Tyrosine Kinases , Genetics
4.
Chinese Journal of Pediatrics ; (12): 851-854, 2012.
Article in Chinese | WPRIM | ID: wpr-348523

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the mutation of IL2RG gene in a Chinese family with a birth history of a dead child suspected of X-linked severe combined immunodeficiency (X-SCID), and to perform prenatal diagnosis with DNA sequencing.</p><p><b>METHOD</b>Blood samples of the parents of the dead child and chorionic villi at gestational age 11 weeks were collected. Eight exons comprising the open reading frame as well as their exon/intron boundaries of IL2RG gene were analyzed by PCR and bi-directional sequencing.</p><p><b>RESULT</b>A heterozygous nucleotide substitution c.690C > T (R226C) in exon 5 was detected in the mother, but not in the father. In the second pregnancy of the mother, the mutation of R226C was not detected in the male fetus by prenatal diagnosis, and the heterozygous mutation was detected in the female fetus of the third pregnancy. The reliability of the prenatal genetic diagnosis was confirmed by the one-year follow-up after the neonates were born.</p><p><b>CONCLUSION</b>The mutation of c.690C>T in IL2RG gene may be the pathologic cause of the proband with X-SCID. DNA sequencing combining sex determination is a valid strategy for prenatal diagnosis of X-SCID.</p>


Subject(s)
Adult , Asians , Genetics , Base Sequence , DNA Mutational Analysis , DNA Primers , Exons , Genetics , Female , Heterozygote , Humans , Infant , Interleukin Receptor Common gamma Subunit , Genetics , Male , Mutation , Pedigree , Polymerase Chain Reaction , Pregnancy , Prenatal Diagnosis , Methods , X-Linked Combined Immunodeficiency Diseases , Diagnosis , Genetics
5.
Chinese Medical Journal ; (24): 2874-2878, 2011.
Article in English | WPRIM | ID: wpr-292786

ABSTRACT

<p><b>BACKGROUND</b>The common γ chain (γc) plays a critical role in regulating proliferation, differentiation, and apoptosis of peripheral T-cells. It was previously confirmed that blocking the γc signal can successfully induce transplant tolerance in a murine model. Here we investigated the potential mechanism.</p><p><b>METHODS</b>Splenocytes from C57BL/6 mice were transfused into T-cell deficient Balb/c nude mice that were reconstituted with syngeneic wild-type T-cells labeled with 5-carboxyfluorescein diacetate succinimidyl ester (CFSE). After 24 hours, recipients received i.p. injection of mixture of anti-γc mAbs, or with isotype control IgG2a. The labeled T-cells were harvested from recipient spleens after 12 and 48 hours. T-cell proliferation and apoptosis were detected by flow cytometry.</p><p><b>RESULTS</b>T-cell proliferation was markedly inhibited and apoptotic T cells could be detected at 12 hours after the mAbs injection. Proliferation was inhibited at 48 hours, but the proportion of apoptotic T-cells was not more than at 12 hours. In the control group, however, T-cells actively proliferated and no significant apoptosis was detected at either time point.</p><p><b>CONCLUSIONS</b>The results suggested that blockade of γc signals can synergize with donor splenocyte transfusion and lead to inhibition of antigen-specific T-cell proliferation and induction of apoptotic T-cell death. This protocol may develop a novel approach to induce donor-specific tolerance.</p>


Subject(s)
Animals , Antibodies, Monoclonal , Pharmacology , Apoptosis , Cells, Cultured , Flow Cytometry , Fluoresceins , Immune Tolerance , Interleukin Receptor Common gamma Subunit , Metabolism , Lymphocyte Activation , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Nude , Signal Transduction , Spleen , Cell Biology , Succinimides , T-Lymphocytes , Cell Biology
6.
Asian Pac J Allergy Immunol ; 2005 Dec; 23(4): 221-6
Article in English | IMSEAR | ID: sea-37065

ABSTRACT

Bacille Calmette-Guerin (BCG) vaccination is used to prevent severe M. tuberculosis infection. It has been used in many countries for a long time. However, complications do occur, including localized abscesses, regional lymphadenitis and disseminated disease. The latter is often associated with underlying immunodeficiency. We report an 8-month-old male infant presenting with cough and fever who had had a generalized pigmented skin rash for one month. Skin biopsy revealed mycobacterial infection, but his response to treatment was poor and he had a persistent mild fever. Immunological studies revealed an IgG of 49 mg/dl, IgA 4 mg/dl, IgM 28 mg/dl, IgE < 1 mg/dl, CD3 1.1%, CD4 0.6%, CD8 0.6%, CD19 93.9%, CD57 1.1%, activated T cells 0.9%, and CH50 < 6.3%. These findings are compatible with the diagnosis of T(-)B(+)NK- severe combined immunodeficiency. Sequence analysis was performed and showed the presence of missense mutation in IL2Rgamma gene. An X-linked recessive inheritance pattern was proved by sequence analysis of his mother and grandmother. In order to identify the strain of the microorganism, we reviewed pathology of the skin biopsy which consisted of diffuse histiocytic infiltrate with poorly formed granulomas and no necrosis and used polymerase chain reaction (PCR) with the stain-positive clinical specimen and verify the organism found in the child's biopsy as M. bovis BCG strain. The diagnosis of disseminated BCG disease must be considered in any infant with cutaneous mycobacterial lesions, especially with atypical histologic findings. Such a patient's immunologic status should be evaluated and further family study is suggested. A high index of suspicion is needed to make a timely diagnosis, as early intervention with intensive treatment and bone marrow transplantation may be life-saving.


Subject(s)
BCG Vaccine/adverse effects , DNA, Bacterial/analysis , Fatal Outcome , Humans , Infant , Interleukin Receptor Common gamma Subunit , Male , Mutation , Mycobacterium Infections/complications , Mycobacterium bovis/genetics , Opportunistic Infections/complications , Receptors, Interleukin/genetics , Severe Combined Immunodeficiency/complications , Skin/pathology , Skin Diseases, Bacterial/complications
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