Impact of transfection with total RNA of K562 cells upon antigen presenting, maturation, and function of human dendritic cells from peripheral blood mononuclear cells.

Background: Vaccination of dendritic cells (DCs) with tumor antigens is a potential strategy to induce tumor-specific immunity in tumor-bearing patients. The purpose of this study was to investigate whether human monocyte-derived DCs were able to present P210(Bcr-Ab1) protein and induce antigen-specific cytotoxic T lymphocyte (CTL) responses in vitro after transfected with total RNA of K562 cells (K562-RNA).

Study Design And Methods: DCs derived from human peripheral blood mononuclear cells were transfected with K562-RNA with electroporation or DOTAP lipofection.

[Proliferation of natural killer T cells in umbilical cord blood and peripheral blood and their different phenotypes].

Purpose of this study was to establish an effective method in vitro to proliferate natural killer T (NKT) cells from umbilical cord blood (UCB) and peripheral blood (PB), and to study their different phenotype. Mononuclear cells (MNC) from UCB and PB were cultured in the presence of IL-2 (100 U/ml), with or without alpha-Galcer. TCR Valpha24 Vbeta11 double positive natural killer T-cells (NKT cells) and their other phenotypes were determined by flow cytometry.

[Experimental study of K562 cell apoptosis induced by siRNA].

Objectives: To construct a siRNA expression vector pBCR6 that produces siRNA against bcr/abl mRNA and detect apoptosis rate of K562 cells after pBCR6 transfection.

Methods: Template sequence for siRNA was designed, synthesized and inserted into an expression vector pSilencer1.0-U6.

[Study on the targeting effects of M1-GS RNA on K562 cells].

Objective: To determine the effects of M1-GS RNA (M1 RNA) on bcr-abl mRNA and oncoprotein after M1 RNA with guide sequence (M1-GS RNA) targeting the oncogene was transfected into K562 cells.

Methods: pAVGS4 (an eukaryocyte expression vector containing M1-GS RNA sequence) and pNAV-1 (as the control) were transfected into K562 cells by X-tremeGENE Q2. Total RNA was extracted at 24, 48, 72 and 96 hours after transfection.

[Inhibition effect of CIITA anti-sense cDNA on the expression of MHCII molecules of Hela cells].

Objective: To investigate the inhibiting effect of classII major histocompatibility complex transactivator (CIITA) anti-sense cDNA on MHCII molecules expression.

Methods: CIITA antisense cDNAs (arII1, arII2 and arII3) were obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid.

[Major histocompatibility complex class II transactivator (CIITA) hammer- head ribozyme transfer inhibits major histocompatibility complex class II (MHC-II) expression in Jukart cells].

Objective: Graft versus host disease (GVHD), a major cause of graft failure in allo-hematopoietic cell transplantation, was associated with the presence of major histocompatibility complex class II (MHCII), also called human leukocyte antigen (HLAII), on the tissues and organs of host. MHCII played a critical role in the induction of immune responses by presenting fragments of allo-antigenic peptides to CD(4)(+) T lymphocytes, then by resulting in CD(8)(+) T lymphocytes activation. Therefore, it was very important for compatibility of MHCII in allo-transplantation.

[The biological activity of MHC classII transactivator ribonuclease P: a novel approach for hepatic transplantation rejection].

Objective: This paper studied the effect of RNaseP against CIITA on repressing class II MHC (MHCII) expression.

Methods: It was constructed that M1-RNA with guide sequences (GS), recognizing the 629 site of CIITA (M1-629-GS), by PCR from pTK117 plasmid, then was cloned into psNAV (psNAV-M1-629-GS). CIITA target gene was obtained from Raji cell by RT-PCR, and then inserted into pGEM-7zf (+) (pGEM-800).

[Cloning and biological activities of riboenzymes against major histocompatibility complex class II transactivator].

Objective: To investigate the cloning and biological activities of riboenzymes against major histocompatibility complex (MHC) class II transactivator (CIITA) so as to explore the feasibility of using hammerhead riboenzyme to create immune tolerance.

Methods: Three riboenzymes against MHC CIITA were synthesized and their activities were evaluated in vitro. Rz464, the riboenzyme with a better digestion effect, was inserted into the vector pIRES2-EGFP (then called pRz464).

Repression of allo-cell transplant rejection through CIITA ribonuclease P+ hepatocyte.

Aim: Allo-cell transplant rejection and autoimmune responses were associated with the presence of class II major histocompatibility complex (MHC II) molecules on cells. This paper studied the effect of Ribonuclease P (RNase P) against CIITA, which was a major regulator of MHCII molecules, on repressing the expression of MHCII molecules on hepatocyte.

Methods: M1-RNA is the catalytic RNA subunit of RNase P from Escherichia coli.