The human leucocyte differentiation antigens (HLDA) workshops: the evolv-ing role of antibodies in research, diagnosis and therapy

The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.

LRRC25 plays a key role in all-trans retinoic acid-induced granulocytic differentiation as a novel potential leukocyte differentiation antigen

Leukocyte differentiation antigens （LDAs） play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 （leucine rich repeat-containing 25） was predicted to have a role in the function of myeloid cells by a large-scale ＂omics＂ data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia （AML） cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid （ATRA）-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia （APL; AML-M3, FAB classification） cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced termi- nal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.

Glomerular chemokine expression and the effect of steroid and cyclophosphamide pulse therapy in human crescentic glomerulonephritis

OBJECTIVE: To study glomerular expression of C-C chemokines, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1alpha and beta (MIP-1alpha, MIP-1beta) and the effect of steroid and cyclophosphamide (CTX) intermittent intravenous pulse therapy on expression in patients with crescentic glomerulonephritis (CGN) to further investigate the underlying mechanism of the treatment. METHODS: Twelve patients with initial biopsy-proven CGN(2), 6 with lupus nephritis (lupus-CGN, LN-CGN) and 6 with vasculitis, (vasculitis-CGN, V-CGN) were enrolled in this study. They underwent an initial biopsy before steroid and CTX intermittent intravenous pulse therapy and were biopsied again one to three months later. Expression of MCP-1, MIP-1alpha, MIP-1beta, and CD68 in glomeruli with cellular and fibrocellular crescents were examined by immunohistochemical analysis in serial sections of renal biopsies. The effect of the pulse therapy on histopathological changes was also observed. RESULTS: Although steroid and CTX intermittent intravenous pulse therapy markedly reduced the degree of glomerular crescent formation both in LN-CGN and V-CGN, the effect of the therapy on glomerular chemokine expression was significantly different between LN-CGN and V-CGN. It was found that steroid and CTX intermittent intravenous pulse therapy reduced the expression of CD68, MCP-1, and MIP-1alpha, but had no effect on MIP-1beta in glomeruli with cellular crescents of patients with LN-CGN. In patients with V-CGN, the therapy also reduced the expression of CD68, but had no effect on MCP-1, MIP-1alpha, and MIP-1beta in glomeruli with cellular crescents. It was noted that the degree of glomerulosclerosis and tubular interstitial fibrosis increased more significantly at the second biopsy in V-CGN as compared to LN-CGN. CONCLUSIONS: The efficacy of steroid and CTX intermittent intravenous pulse therapy in CGN might be affected by reduction of glomerular chemokine expression. The different changes in glomerular expression of MCP-1 and MIP-1alpha in patients with LN-CGN and V-CGN after pulse therapy may correlate to different responses to treatment and prognosis.

CTLA-4 gene A/G polymorphism associated with diabetes mellitus in Han Chinese

OBJECTIVE: To investigate the association of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene A/G polymorphism with susceptibility to diabetes mellitus in Han Chinese. METHODS: An A/G transition at position 49 of exon 1 was analyzed in 31 patients with type 1 diabetes, 31 patients with type 2 diabetes, and 36 controls were analyzed by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: A highly significant increase in the frequency of the G allele was seen in patients with type 1 diabetes compared with controls (66.1 % vs. 34.7%, respectively; P

Clinical observation of Yiqi Qingdu Prescription(益气清毒方)on the treatment of intermediate-stage and advanced non-small-cell lung cancer

OBJECTIVE:To observe the effects of the Yiqi Qingdu prescription(益气清毒方)on intermediate-stage and advanced non-small-cell lung cancer(NSCLC).METHODS:In total,300 patients with intermediate-stage or advanced NSCLC were randomly and equally divided into three groups using computer-generated random numbers as follows:Western medicine(WM),Chinese medicine(CM),and integrated Traditional Chinese and Western Medicine(IM).After 3 months of treatment,the overall response rate(ORR);disease control rate(DCR);symptom score(SS);Karnofsky performance status(KPS);adverse event score;counts of CD3^(+),CD4^(+),and CD8^(+)cells;CD4^(+)/CD8^(+)ratio;and carcinoembryonic antigen(CEA)level were compared among the groups.RESULTS:The ORRs were 30.36%,20.24%,and 7.87%in the IM,CM,and WM groups,respectively,whereas the DCRs were 85%,75%,and 73%,respectively.Compared to the CM group,the ORR was significantly higher in the WM and IM groups,whereas the DCR was significantly higher in the IM group(all P<0.05).SS was obviously higher in the WM group than in the other two groups(both P<0.01).KPS was significantly lower in the WM group after treatment(P=0.005).The mean number of adverse events was significantly lower in the CM(2.2±1.3)and IM(2.4±1.3)groups than in the WM group(4.6±1.7,both P<0.05).CD3^(+)cell counts were significantly decreased in the WM group(P=0.031).In the IM group,CD8^(+)cell counts were increased after treatment,whereas the CD4^(+)/CD8^(+)ratio was decreased(both P<0.01).Compared with the WM group,CD3^(+)(P=0.01),CD4^(+)(P=0.044),and CD8^(+)(P=0.009)cell counts were significantly higher in the IM group,whereas the CD4^(+)/CD8^(+)ratio was significantly lower(P=0.011).Relative to the CM group,CD8^(+)cell counts were significantly higher(P=0.001)and the CD4^(+)/CD8^(+)ratio was significant ly lower in the IM group(P=0.001).CEA levels were significantly increased in the CM group(P=0.023).CONCLUSION:The Yiqi Qingdu prescription can improve the outcomes of WM in patients with NSCLC.

Immunophenotype of myeloid cells in myelodysplastic syndromes and its clinical implications

OBJECTIVE: To explore the immunophenotype of myeloid cells in myelodysplastic syndyomes (MDS) and its clinical implications. METHODS: A panel of monoclonal antibody was used to detect CD13+, CD33+, CD15+ and CD14+ antigens on the membrane surfaces of myeloid cells in the bone marrow from 51 MDS, 21 aplastic anemia (AA), 21 paroxysmal nocturnal hemoglobinuria (PNH) patients. 10 acute myeloblastic leukemia (AML) patients and 15 normal subjects by immunoenzymatic assay. The morphology and chromosome karyotype of bone marrow cells of MDS patients were also examined. RESULTS: CD14+, CD13+ and CD33+ cells in the bone marrow were more in MDS patients than in normal controls, AA patients and PNH patients. CD15+ cells in the bone marrow were less in MDS patients than in normal controls. CONCLUSIONS: The percentages of CD14+, CD13+ and CD33+ positive cells in the bone marrow of MDS patients were related to the percentage of myeloblasts, the chromosomal aberrations and the response to treatment. It indicated that there is immunophenotypic misexpression of myeloid cells in MDS patients. Immunophenotype analysis of myeloid cells might be useful for the diagnosis and treatment of MDS patients.