[Preclinical study of T cell receptor specifically reactive with KRAS G12V mutation in the treatment of malignant tumors].

OBJECTIVE: KRAS gene is one of the most common mutations of proto-oncogenes in human tumors, G12V is one of the most common mutation types for KRAS. It's challenging to chemically acquire the targeted drug for this mutation. Recent studies reported that this mutation peptides can form a neoepitope for T cell recognition. Our study aims to clone the T cell receptor (TCR) which specifically recognizes the neoepitope for KRAS G12V mutation and constructs TCR engineered T cells (TCR-T), and to investigate if TCR-Ts have strong antitumor response ability. METHODS: In this study, tumor infiltrating lymphocytes were obtained from one colorectal cancer patient carrying KRAS G12V mutation. Tumor-reactive TCR was obtained by single-cell RT-5' rapid-amplification of cDNA ends PCR analysis and introduced into peripheral blood lymphocytes to generate TCR-Ts. RESULTS: We obtained a high-affinity TCR sequence that specifically recognized the HLA-A*11:01-restricted KRAS G12V8-16 epitope: KVA11-01. KVA11-01 TCR-T could significantly kill various tumor cells such as PANC-1, SW480 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V), and secreting high levels of interferon-γ (IFN-γ). Non-specific killing experiments suggested KVA11-01 specifically recognized tumor cells expressing both mutant KRAS G12V and HLA-A*11:01. In vivo assay, tumor inhibition experiments demonstrated that infusion of approximately 1E7 KVA11-01 TCR-T could significantly inhibit the growth of subcuta-neously transplanted tumors of PANC-1 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V) cells in nude mice. No destruction of the morphologies of the liver, spleen and brain were observed. We also found that KVA11-01 TCR-T could significantly infiltrate into tumor tissue and had a better homing ability. CONCLUSION: KVA11-01 TCR-T cells can effectively target a variety of malignant tumor cells carrying KRAS G12V mutation through in vitro and in vivo assay. KVA11-01 TCR-T cells have excellent biological activity, high specificity of target antigen and homing ability into solid tumor tissue. KVA11-01 TCR-T is expected to be an effective treatment for patients with KRAS G12V mutant solid malignancies.

Research on the diagnostic effect of PCT level in serum on patients with sepsis due to different pathogenic causes.

OBJECTIVE: To investigate the diagnostic effect of procalcitonin level in serum for patients with sepsis due to different pathogenic causes. PATIENTS AND METHODS: The clinical data of 132 sepsis patients were analyzed. Those patients were admitted to the Affiliated Hospital of Medical School of Ningbo University from January 2014 to January 2017. According to the blood culture results before antimicrobial therapy, patients were divided into two groups: Gram-negative bacteria group (G- group) and Gram-positive bacteria group (G+ group). The indexes, such as SOFA score, APACHE II score, length of stay in hospital and mortality rate, were used to evaluate disease severity of the two groups. The procalcitonin, WBC, hs-CRP and NEU% were detected and compared between the two groups of patients. RESULTS: A total of 132 pathogenic bacteria were detected in 132 patients, of which 44 patients were infected with G- bacteria and 88 patients were infected with G+ bacteria. Patients in G- group were mainly infected with Escherichia coli, Acinetobacter baumannii, and Klebsiella pneumoniae, while patients in G+ group were mainly infected with Staphylococcus epidermidis and Staphylococcus aureus. The SOFA score, APACHE II score and mortality rate in G- group were higher than those in G+ group. The PCT levels in G- group and G+ group were (54.89±21.64) ng/mL and (21.13±1.30) ng/mL, respectively. The PCT level in G- group was higher than that in G+ group, and the difference was statistically significant between them (p<0.05). There was no statistically significant difference in length of stay in hospital between the two groups (p>0.05). There were no statistically significant differences in WBC, hs-CRP and NEU% between the two groups (p>0.05). CONCLUSIONS: The procalcitonin level in serum of sepsis patients at early stage of bloodstream infection is significantly elevated and has diagnostic value for different pathogenic bacteria groups. It can also reflect the disease severity and predict the prognosis of sepsis patients.

Oral testosterone undecanoate (Andriol) supplement therapy improves the quality of life for men with testosterone deficiency.

In a single-blind, placebo-controlled study, the effects of a 3-month oral administration of 160 mg/day testosterone undecanoate (Andriol) on the quality of life of men with testosterone deficiency were evaluated. The subjects included ten men with primary hypogonadism and 29 with andropause with sexual dysfunction as the most common problem. The changes in subjective symptoms were evaluated by the PNUH QoL scoring system and the St. Louis University Questionnaire for androgen deficiency in aging males (ADAM). Digital rectal examination (DRE) was performed and serum testosterone, prostate-specific antigen (PSA) and liver profile were monitored. Testosterone undecanoate treatment (n = 33) significantly improved sexual dysfunction and symptom scores of metabolic, cardiopulmonary, musculoskeletal and gastrointestinal functions compared to baseline and to placebo (n = 6). ADAM score also significantly improved after 3 months of treatment. Serum testosterone was significantly increased compared to pretreatment levels only in the testosterone undecanoate group. In the placebo group, no significant changes compared to baseline were found for testosterone levels and QoL questionnaires. No abnormal findings were detected on DRE or laboratory findings in either group. Adverse events, such as gastrointestinal problems and fatigue, were mild and self-limiting. It is concluded that androgen supplement therapy with oral testosterone undecanoate (Andriol) restores the quality of life through improvement of general body functions in men with testosterone deficiency.