Inducement of Specific CTLs by Antigen-Peptides from Human Leukemia Cells and Their Cytotoxicity to Leukemia Cells

To investigate the inducement of cytotoxic T lym phocytes(CTL s) by antigen peptides m ixture from different leukemia cells and the cross- reaction of the m ixtures from different cell lines,antigen peptides m ixtures were prepared from different leukemia cell lines respectively and then bound with Hsp70 in vitro.Activation and proliferation of PBMC were observed after stim u- lation with different Hsp70 - peptide complexes.The ratio of CD8+ in proliferative cells was ana- lyzed by flow cytometry.The cytotoxicity of the activated PBMC to different target cells was as- sayed.The results showed that the antigen peptides from different leukemia cell lines,bound with Hsp70 ,could activate PBMC effectively,and stimulate the activated PBMC to proliferate.The proliferative PBMC had specific cytotoxicity to corresponding leukem ia cells.CD8+ cells,account- ing for a high proportion in proliferative cells,had a specific cytotoxicity to leukemia cells from which antigen peptides were prepared,suggesting that these CD8+ cells were CTL s specific to leukemia cells.CTL s activated by Hut78- peptides or Molt4 - peptides had a significantly stronger cytotoxicity to Hut78cells,Molt4 cells and Jurkat cells than that of CTL s activated by HL - 6 0 - peptides(P<0 .0 5 ) .And the cytotoxicity of CTL s activated by Hut78/ Molt4 - peptides to Jurkat cells was significantly stronger than that of CTL s activated by either Hut78- peptides or Molt4 - peptides alone(P<0 .0 5 ) .It is concluded that antigen peptides m ixtures from leukem ia cells can induce specific antitumor CTL s.There exists cross- reactivity among antigen peptides m ixtures from different cell lines of the sam e type leukemia and more cross- reactive antigen peptides could be obtained from m ore cell lines,suggesting that antigen peptides m ixture with broad antigenic spectrum could be prepared by using multiple leukemia cell lines.

Prediction, Synthesis and Antigenicity of the Antigenic Peptides of 26kDa Glutathione S-Transferasc of Schistosoma Japonicum (Sj26)

Six antigenic peptides of 26 kDa glutathione S-transferase of Schistosoma japonicum(Sj26) have been predicted according to their hydrophilicity, flexibility. accessibility. chargedistribution and β -turn in the secondary structure by the determination of its primary structure andsynthesized by solid phase method. All of them showed antigenicity with anti-schistosomajaponicum immunoglobulin polyclonal antibody, anti-Sj-lgG PcAb by Dot-ELISA. Three of themshowed good antigenicity. They would serve as candidates of synthetic anti-schistosomal vaccine.

Mapping the Antigenic Peptides of Sm26/2 in Glutathione S-Transferases of Schistosoma Mansoni

Six antigenic peptides of Sm26/2 glutathione S-transferase of schistosoma mansoni have been predicted according to their hydrophilicity, flexibility, accessibility, charge distribution and beta-turn in the secondary structure by the determination of its primary structure, and synthesized by solid phase method. Two of them showed good antigenicity by Dot-ELISA. They would be candidate peptides of synthetic anti-schistosomal vaccine.

INVESTIGATION OF INDUCING EFFECT OF SPECIFIC CYTOTOXICITY OF CTLS BY ANTIGEN PEPTIDES FROM T LYMPHOCYTIC LEUKEMIA CELLS

Objective: To investigate the characteristics of specific antitumor immunity induced by antigen peptides mixture from T lymphocytic leukemia cells. Method: Antigen peptides mixtures were prepared from different leukemia cell lines and then bound with Hsp70 in vitro. Human peripheral blood mononuclear cells (PBMC) were cultured in vitro, and activated with Hsp70-antigen peptides. The activated PBMC was cultured continuously in vitro, and used as effector cells in vitro test of cytotoxicity to different target cells. Results: The antigen peptides from different leukemia cell lines were peptides mixture and could activate PBMC effectively if they were presented by Hsp70. The activated PBMC could proliferate in the presence of IL-2 and Hsp70-antigen peptides. The proliferative PBMC had specific cytotoxicity to leukemia cells corresponding to the antigen peptides. PBMC activated by antigen peptides from T lymphocytic leukemia cell lines could effectively kill T lymphocytic leukemia cells, and the cytotoxicity of these PBMC to T lymphocytic leukemia cells was significantly stronger than that of PBMC activated by antigen peptides from other leukemia cells (P < 0.05). PBMC activated by either Hut78-peptides or Molt 4-peptides could effectively kill Jurkat cells. And the cytotoxicity of PBMC activated by Hut78/Molt-4-peptides to Jurkat cells was significantly stronger than that of PBMC activated by either Hut78-peptides or Molt-4-peptides alone (P<0.05). Conclusion: Antigen peptides mixture from T lymphocytic leukemia cell lines can induce specific cytotoxic effect to T lymphocytic leukemia cells. There exists cross-reactivity among antigen peptides mixture from different T lymphocytic leukemia cell lines. The cross-reactivity could be amplified by blending of different antigen peptides from different T lymphocytic leukemia cell lines, suggesting that it is possible to prepare broad-spectrum antigen peptide vaccine against T lymphocytic leukemia by using multiple leukemia cell lines.

Advances in Structure-function Relationships and Mechanism of Transporter Associated with Antigen Processing

The transporter associated with antigen processing （TAP） belongs to the ATP-binding cassette （ABC） transporter superfamily. Driven by ATP hydrolysis, TAP translocates antigenic peptides from the cytosol into the ER （endoplasmic reticulum） lumen where the antigenic peptides are loaded onto the HLA class I molecules. Recently, numerous studies show that TAP is closely related with various diseases such as viral infections, autoimmune diseases, and different malignancies. In consideration of important roles of TAP in human adaptive immunity, this review summarizes the recent advances in structure-function relationships of crucial domains and transport mechanism systematically. The challenging problems and potential methods are also pointed out for in-depth researches.

Study on the Preparation of Recombinant Human HSP70 and Its Presenting-Antigen Function

The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in glucose free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol/L and 5 mmol/L respectively, the engineered bacteria carrying expression vector of human HSP70 gene expressed rHSP70 at an efficiency of 60 %. After the purification with DEAE ion exchange chromatography, HSP70 with a purity of higher than 90 % was obtained. The purified product could bind tumor antigen peptide in vitro , and the binding was identified by native PAGE containing 5 % glycerol. HSP70 peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells, suggesting that the recombinant human HSP70 could be used as an antigen presenting reagent in tumor therapy.

Antigenic Peptide Loading into Major Histocompatibility Complex Class I Is Driven by the Substrate N-Terminus

Major histocompatibility complex class I(MHC-I),a key element of the acquired immune system,plays essential roles in activating CD8^(+)T cells by recognizing intracellular antigens derived from pathogens and cancer.Assembly of MHC-I and antigen peptides is critical for the antigen presentation on the cell surface.However,the structural dynamics of antigenic peptide loading into MHC-I,at atomistic resolution,is still elusive.Here,by constructing a Markov state model(MSM)based onlarge scale all-atommolecular dynamics(MDs)simulations with an aggregated simulation time∼24μs,we reveal the detailed molecular mechanism underlying the peptide-loading dynamics into MHC-I and identify the key intermediates with associated thermodynamic/kinetic properties.Furthermore,we examine how the chaperone tapasin-binding protein related(TAPBPR)participates in promoting the peptide loading,and the results show that TAPBPR,by binding to the F pocket,allosterically modulates the structures of the distant pocket B,resulting in formation of a peptide-receptive conformation ideal for accommodating the incoming peptide N-terminus.This study provides fundamental structural insights for the peptide loading into MHC-I in both chaperone uncatalyzed and catalyzed contexts.

The dichotomous role of immunoproteasome in cancer:Friend or foe?

Immunoproteasome is a variant of proteasome with structural differences in 20S subunits optimizing them for the production of antigenic peptides with higher binding affinity to major histocompatibility complex(MHC)-I molecules.Apart from this primary function in antigen presentation,immunoproteasome is also responsible for the degradation of proteins,both unfolded proteins for the maintenance of protein homeostasis and tumor suppressor proteins contributing to tumor progression.The altered expression of immunoproteasome is frequently observed in cancers;however,its expression levels and effects vary among different cancer types exhibiting antagonistic roles in tumor development.This review focuses on the dichotomous role of immunoproteasome in different cancer types,as well as summarizes the current progression in immunoproteasome activators and inhibitors.Specifically targeting immunoproteasome may be a beneficial therapeutic intervention in cancer treatment and understanding the role of immunoproteasome in cancers will provide a significant therapeutic insight for the prevention and treatment of cancers.