IgG subclass distribution of anti-Rh,anti-Kell and anti-Duffy antibodies measured by sensitive haemagglutination assays.

The IgG subclass distribution of anti Rh antibodies (anti-D, ''anti-Du'', anti-c, anti-E), anti-Kell and anti-Duffy (anti-Fya) antibodies was measured by two haemagglutination techniques on microtitre plates. The first technique involved rabbit subclass specific antisera which were used to agglutinate red cells previously reacted with the patients'' antibodies at high concentration. The second, which was more sensitive, had an additional step by introducing sheep anti-rabbit antibodies (sandwich technique). By the sensitive sandwich technique we revealed, for anti-D antibodies: IgG1 8/19, IgG3 1/19, IgG1/IgG3 8/19, IgG1/IgG2/IgG3/IgG 41/19, IgG1/IgG4 1/19; for the Du reactive anti-D antibodies: IgG1 1/8, IgG1/IgG3 1/8, IgG1/IgG3/IgG4 6/8; for the anti-E antibodies: IgG1/IgG2/IgG4 2/3, IgG1/IgG2/IgG3/IgG4 1/3; for the anti-c antibodies: IgG1 2/5, IgG3 1/5, IgG1/IgG3 1/5; for the anti-Kell antibodies: IgG1 9/20, IgG1/IgG3 1/20, IgG1/IgG4 8/20, IgG1/IgG3/IgG4 2/20; and for anti-Duffy antibodies: IgG1 1/8, IgG1/IgG4 7/8. These results are partly at variance with previously published results.     

The production and characterization of two monoclonal antibodies to rat kidney L-arginine: glycine amidinotransferase

Rat kidney L-arginine:glycine amidinotransferase (transamidinase) has been purified previously to homogeneity as two fractions, designated alpha and beta. No differences in the properties of these two fractions could be found. Two monoclonal antibodies (Tran/NS-1/1 and Tran/NS-1/3) to the purified alpha fraction of rat kidney transamidinase were produced, purified, and characterized. The results of competitive binding studies of the two monoclonal antibodies to alpha transamidinase were as follows: 1) Tran/NS-1/3 had no effect on 125I-Tran/NS-1/1 binding while Tran/NS-1/1 inhibited 125I-Tran/NS-1/1 binding; 2) Tran/NS-1/3 inhibited 125I-Tran/NS-1/3 binding while Tran/NS-1/1 had no effect on 125I-Tran/NS-1/3 binding. Therefore, Tran/NS-1/1 and Tran/NS-1/3 bound to different antigenic determinants on alpha transamidinase. 125I-Tran/NS-1/1 and 125I-Tran/NS-1/3 each had high avidity constants (approximately 10(7)-10(9)) for both alpha and beta rat kidney transamidinase. Tran/NS-1/1 and Tran/NS-1/3 bound to human kidney transamidinase in ELISA assays. A quantitative immunosorbent inhibition assay for rat kidney transamidinase was developed with 125I-Tran/NS-1/3. Approximately 30 ng of immunoreactive transamidinase could be detected by this immunosorbent inhibition assay. The amount of Tran/NS-1/3 immunoreactive species in rat lung and testicular tissue by the immunosorbent inhibition assay correlated well with the amount of transamidinase activity found in those tissues. The availability of the monoclonal antibodies, Tran/NS-1/1 and Tran/NS-1/3, should facilitate studies of rat and human transamidinase structure and regulation.     

Expression and Subcellular Localization of Recombinant SDF-1 and Its Mutant Intrakine in Transfected COS-7 Cells

INTRODUCTION The CXC chemokine stromal cell -derived factor1(SDF-1) , a member of CXC-chemokine family,is the ligand for CXCR4 and binds CXCR4 with high affinity.The interaction of SDF-1 and CXCR4 mayinduce cytoskeletal rearrangement, adhesive to endothelial cells and directional migration. SDF-1 bind-ing to CXCR4 can activiate multiple signaling pathways and produce different physiology or pathologyeffects. Compelling evidence is accumulating HIV infection and tumor metastas…     

Palmitoylation of the sphingosine 1-phosphate receptor S1P1 is involved in its signaling functions and internalization

The lipid mediator sphingosine 1-phosphate (S1P) regulates several cellular processes through binding to its receptors (S1P₁–S1P₅), which are heterotrimeric G protein-coupled receptors. Here, we report that all S1P receptors are palmitoylated. In S1P₁, three Cys residues in the cytoplasmic tail are palmitoylated. We examined the roles of palmitoylation of S1P₁ using model cells in which wild-type S1P₁ or a non-palmitoylated mutant S1P₁ was overproduced. Compared with wild-type S1P₁, the non-palmitoylated S1P₁ exhibited binding affinity similar to the natural ligand S1P but lower to the synthetic ligand FTY720 phosphate (FTY720-P), the active form of the immunomodulator FTY720. However, downstream signaling of non-palmitoylated S1P₁ was similarly affected by S1P and FTY720-P stimulation. Moreover, upon stimulation with S1P, internalization of the mutant non-palmitoylated S1P₁ was retarded, compared with that of the wild-type protein. This effect was much more pronounced with FTY720-P stimulation. Similar differences were observed for the phosphorylation of S1P₁ and its mutant. These findings may provide insights into the molecular mechanisms of the pharmacological effects of FTY720. Finally, palmitoylation of wild-type S1P₁ increased upon treatment with S1P, suggesting that S1P₁ undergoes a palmitoylation/depalmitoylation cycle after stimulation by its ligands.     

Further evidence for infection of pigs with human-like H1N1 influenza viruses in China

Classical swine and avian-like H1N1 influenza viruses were reported widely in swine population worldwide, but human-like H1N1 swine viruses were reported occasionally. In 2006, a human-like H1N1 swine virus (A/swine/Guangdong/96/06) was isolated from pigs in Guangdong province, which was reported in China for the first time. To get further evidence for infection of pigs with human-like H1N1 influenza viruses, we analyzed eight gene segments of three human-like swine H1N1 viruses (A/swine/Guangdong/96/06, A/swine/Tianjin/01/04 and A/swine/Henan/01/06) isolated in China. All the eight genes of the three viruses are highly homologous to recent (about 2000) and early (1980s) human H1N1 influenza viruses, respectively. Phylogenetic analyses revealed that A/Swine/Guangdong/96/06 was directly derived from about 2000 human H1N1 influenza viruses, while A/swine/Tianjin/01/04 and A/swine/Henan/01/06 seemed to be descendants of human H1N1 viruses circulating in 1980s. Seroprevalence of our isolate (A/swine/Guangdong/96/06) confirmed the presence of human-like H1N1 virus in pigs in China. Existence of these influenza viruses, especially older viruses (A/swine/Tianjin/01/04 and A/swine/Henan/01/06), indicates that human-like H1N1 influenza viruses may remain invariant for long periods in pigs and provides the evidence that pigs serve as reservoirs of older influenza viruses for human pandemics.     

A/PR/8/34(H1N1)病毒株血清流行病学调查

目的了解当今人群中是否仍有Ａ／ＰＲ／８／３４（Ｈ１Ｎ１）这类毒株的活动。方法从全国８个不同地区采集１９７５年之后出生人的血清，用常量半加敏ＨＩ测定Ａ／ＰＲ／８／３４（Ｈ１Ｎ１）毒株的抗体，然后凡ＨＩ效价≥１０的血清，一律按同法对Ａ／Ｓｗｉｎｅ／Ｉｏｗａ／１５／３０（Ｈ１Ｎ１）、Ａ／桂富／１０／１９４（Ｈ１Ｎ１）和Ａ／京防／５３／９７（Ｈ１Ｎ１）毒株进行复核测定。结果有２４份血清ＨＩ效价≥２０，分布于８个不同地区。结论当今我国人群中存在有Ａ／ＰＲ／８／３４（Ｈ１Ｎ１）毒株的活动     

Contribution of IL-1 to resistance to Streptococcus pneumoniae infection

The role of IL-1 in susceptibility to Streptococcus pneumoniae infection was studied in mice deficient in genes of the IL-1 family [i.e. IL-1alpha-/-, IL-1beta-/-, IL-1alpha/beta-/- and IL-1R antagonist (IL-1Ra)-/- mice] following intra-nasal inoculation. Intra-nasal inoculation of S. pneumoniae of IL-1beta-/- and IL-1alpha/beta-/- mice displayed significantly lower survival rates and higher nasopharyngeal and lung bacterial load as compared with control, IL-1alpha-/- and IL-1Ra-/- mice. Treatment of IL-1beta-/- mice with rIL-1beta significantly improved their survival. A significant increase in blood neutrophils was found in control, IL-1alpha-/- and IL-1Ra-/- but not in IL-1beta-/- and IL-1alpha/beta-/- mice. Local infiltrates of neutrophils and relatively preserved organ architecture were observed in the lungs of IL-1alpha-/- and control mice. However, S. pneumoniae-infected IL-1beta-/-, IL-1alpha/beta-/- and IL-1Ra-/- mice demonstrated diffuse pneumonia and tissue damage. Altogether, all three isoforms contribute to protection against S. pneumoniae; our results point to differential role of IL-1alpha and IL-1beta in the pathogenesis and control of S. pneumoniae infection and suggest that IL-1beta has a major role in resistance to primary pneumococcal infection while the role of IL-1alpha is less important.     

Monoclonal antibodies to the haemagglutinin HA1 subunit of the pandemic influenza A/H1N1 2009 virus and potential application to serodiagnosis

In order to provide specific serological reagents for pandemic influenza A/H1N1 2009 virus, monoclonal antibodies (Mabs) to recombinant haemagglutinin component HA1 (rHA1) were generated after fusing spleen cells from a mouse immunized with rHA1 protein derived from influenza strain A/California/06/09 H1N1 with a mouse myeloma cell line. Five hybridoma clones secreting Mabs specific for the rHA1 protein derived from pandemic influenza A/H1N1 2009 and not for rHA1 from seasonal H1N1 influenza strains A/Brisbane/59/07 and A/Solomon Islands/03/06 were identified by EIA. Mabs 7H4, 9A4, and 9E12 were reactive in Western blots with full length rHA and/or rHA1 subunit derived from A/California/06/09 strain. Only Mab 1F5 inhibited haemagglutination of turkey red blood cells with recombinant NIBRG‐121 virus derived from A/California/07/09, but did not react in Western blots. Immunostaining of MDCK cells infected with NIBRG‐121 was localized to the membrane/cytoplasm for four of the reactive Mabs. The differing reactivity of the Mabs in Western blots, immunostaining, EIA, and haemagglutination inhibition assay suggest that at least four of the five Mabs recognize different epitopes on HA1 of the pandemic influenza A/H1N1 2009 virus. Ferret antisera to pandemic influenza A/H1N1 2009 (A/England/195/09 and A/California/07/09 strains) and sera from human subjects vaccinated with Influenza A (H1N1) 2009 Monovalent Vaccine (CELTURA®, Novartis Vaccines, Germany), inhibit binding of 1F5‐HRP to biotinylated rHA1 derived from A/California/06/09 in a competitive EIA, suggesting that the epitope recognized by this Mab also evokes an antibody response in infected ferrets and vaccinated humans. J. Med. Virol. 83:559–567, 2011. © 2011 Wiley‐Liss, Inc.     

Enhanced susceptibility to immune nephritis in DBA/1 mice is contingent upon IL-1 expression

The DBA/1 mouse strain is particularly sensitive to experimental immune-mediated nephritis. Previous studies have indicated that various chemokines/cytokines are elevated in strains of mice susceptible to immune-mediated glomerulonephritis. One of the many elevated cytokines is IL-1. This study was designed to determine if IL-1 is essential for the development of immune-mediated nephritis in the DBA/1 mouse strain that is particularly sensitive to this disease. Both male and female DBA/1 mice and DBA/1.IL-1R1(-/-) mice were challenged with anti-GBM sera. We then compared DBA/1 mice to DBA/1.IL-1R1(-/-) mice to determine the influence of IL-1 on immune-mediated nephritis. Compared to DBA/1 mice, DBA/1.IL-1R1(-/-) mice excreted significantly less protein post anti-GBM serum challenge. None of the DBA/1.IL-1R1(-/-) mice, male or female, had a BUN higher than 22 mg/dl post-challenge whereas wild-type DBA/1 mice had significantly elevated BUN. Wild-type DBA/1 mice exhibited pronounced glomerulonephritis, with crescent formation, as well as tubulo-interstitial disease compared to DBA/1.IL1R1(-/-) mice. These findings indicate that IL-1 is necessary for the development of nephritis in DBA/1 mice and suggest that the elevated IL-1 levels in these mice may be one reason why the DBA/1 strain is particularly sensitive to multiple end organ diseases.     

Regulation of Thy-1 Gene Expression by the Methylation of the 5' Region of Thy-1 Gene and Intracellular Regulatory Factors in Immature B Cells

In cμ^- B220⁺ Thy-1^- FL2-52-2, an immature B cell line transformed with a temperature-sensitive mutant of Abelson murine leukemia virus(ts OS-59), Thy-1 antigen expression was induced after the shift of the culture temperature from a permissive (35' C) to a non-permissive temperature (39' C), and a Thy-1⁺ subclone, FL2-52-2-1 was isolated by limiting dilution. Furthermore, since a population of Thy-1⁺FL2-52-2-1 lost Thy-1 antigen expression during culture at a non-permissive temperature, Thy-1^- FL2-52-2-1-1 was isolated from the cultured cells. Methylation analysis by Southern blotting experiments showed that 5' region of the Thy-1 gene was methylated in Thy-1^- FL2-52-2 but demethylated in Thy-1⁺ FL2-52-2-1 and Thy-1^- FL2-52-2-1-1. To determine whether or not there exist intracellular regulatory factors responsible for Thy-1 gene expression, Thy-1^- allele was transfected into Thy-1^- (Thy-1.2^-) FL2-52-2 and Thy-1^- FL2-52-2-1-1. The transfected Thy-1^- allele was expressed in Thy-1^- FL2-52-2, but not in Thy-1^- FL2-52-2-1-1, indicating the presence of the intracellular regulatory factors requisite for Thy-1 gene expression in Thy-1^- FL2-52-2. It appeared that in Thy-1^- FL2-52-2-1-1, Thy-1 gene was not expressed because of the abscence of the intracellular regulatory factors although the 5' region of the Thy-1 gene was demethylated. These results indicated the existence of at least two regulatory mechanisms of Thy-1 gene expression in immature B cells: methylation of the 5' region of the Thy-1 gene and intracellular regulatory factors.     

Two genotypes of H1N2 swine influenza viruses appeared among pigs in China

BackgroundH1N2 is one of the main subtypes of influenza, which circulates in swine all over the world.ObjectivesTo investigate the prevalence and genetic of H1N2 in swine of China.Study designTwo H1N2 swine influenza viruses were isolated from Tianjin and Guangdong province of China in 2004 and 2006, respectively. The molecular evolution of eight gene segments was analyzed.ResultA/Swine/Tianjin/1/2004 has low identity with A/Swine/Guangdong/2006; in the phylogenetic tree of PA gene, A/Swine/Guangdong/1/2006 and A/Swine/Guangxi/1/2006 along with the H1N2 swine isolates of North America formed a cluster; and A/Swine/Tianjin/2004 and A/Swine/Zhejiang/2004, along with the classical H1N1 swine isolates formed another cluster; except that NA gene of A/Swine/Tianjin/1/2004 fell into the cluster of the H3N2 human influenza virus, indicating the reassortment between H3N2 human and H1N1 swine influenza viruses.ConclusionTwo different genotypes of H1N2 appeared among pigs in China. A/swine/Guangdong/1/06 was probably from H1N2 swine influenza viruses of North America; while A/swine/Tianjin/1/04 maybe come from reassortments of classical H1N1 swine and H3N2 human viruses prevalent in North America.     

从人群中分离到1株A／PR／8／34（H1N1）类病毒

１９９１年５月从１名上呼吸道感染的儿童患者中分离出１株流感病毒。经血清学鉴定和分析，分离物为Ａ／ＰＲ／８／３４（Ｈ１Ｎ１）类似毒株。病毒ＨＡ１区基因经核苷酸序列分析，分离物与Ａ／ＰＲ／８／３４毒株间有１２个位点不同，导致了ＨＡ１区蛋白分子上４个氨基酸的替换。在病毒基因组寡核苷酸指纹图分析中发现，分离物与Ａ／ＰＲ／８／３４毒株相比较丢失了２个点，而插入了９个点。因此，Ａ／广东／６／９１（Ｈ１Ｎ１）毒株不是由Ａ／ＰＲ／８／３４（Ｈ１Ｎ１）通过实验室污染而来。     

Evolution of influenza polymerase: nucleotide sequence of the PB2 gene of A/Chile/1/83 (H1N1)

Summary The complete nucleotide sequence of the PB2 gene of influenza virus A/Chile/1/83 (H1N1) is presented. Sequence comparison between A/Chile PB2 protein and the known PB2 sequences of the influenza strains A/WSN/33 (H1N1), A/PR/8/34 (H1N1), A/NT/60/68 (H3N2), A/Kiev/59/79 (H1N1), A/FPV/Rostock/34 (H7N1), and B/Ann Arbor/1/66 indicates extensive amino acid homology for the influenza A virus PB2 proteins. Small clusters of basic amino acids are conserved in all PB2 proteins including the influenza B PB2 protein which has only 39% sequence homology overall to the PB2 polypeptides of type A influenza viruses. The evolutionary rate of 5.7 × 10^–3 nucleotide substitutions per site per year and 0.25% amino acid changes per year between the A/Chile/1/83 and A/NT/60/68 PB2 appears to be higher than that calculated earlier for A/NT, A/PR/8 and A/WSN. An unusually high degree of sequence change between A/Chile/1/83 and A/Kiev/59/79 PB2 polymerase was revealed and this is discussed in terms of its probable origin.     

Comparison of pro-inflammatory cytokine expression and cellular signal transduction in human macrophages infected with different influenza A viruses

Influenza A virus infection of macrophages and virus-induced pro-inflammatory gene expression are regarded to contribute to severity of influenza A virus-caused diseases. Although some data are available on cytokine production by influenza A virus-infected macrophages, systematic comparisons of the virus types are currently considered to be of high relevance in humans (pandemic H1N1/2009, seasonal H1N1, seasonal H3N2, highly pathogenic avian influenza H5N1) on pro-inflammatory potential, and relevant underlying cellular signalling events are missing. Here, we show that the infection of human monocyte-derived macrophages with pandemic H1N1/2009 (A/HH/01/2009), seasonal H1N1/1999 (A/New Caledonia/20/99), seasonal H3N2/2004 (A/California/7/2004) or highly pathogenic H5N1/2004 (A/Thailand/1(Kan-1)/04) results in similar infection rates. However, the investigated H1N1 strains caused delayed and decreased apoptosis in comparison with H3N2/2004 or H5N1/2004. Moreover, human macrophage infection with H3N2/2004 or H5N1/2004 but not with H1N1 viruses was associated with pronounced pro-inflammatory cytokine production and activation of relevant mitogen-activated protein kinase pathways as indicated by phosphorylation of p38, JNK and ERK 1/2. These findings are in line with clinical observations indicating enhanced disease severity in H3N2- or H5N1-infected patients compared to individuals infected with pandemic H1N1/2009 or seasonal H1N1.     

早衰蛋白-1基因内含子多态性与Alzheimer's病的关系

为探讨早衰蛋白－１（ＰＳ－１）基因多态性与Ａｌｚｈｅｉｍｅｒ’ｓ病（ＡＤ０的关系,用聚合酶链反应－限制性片段长度多态性（ＰＣＲ－ＲＦＬＰ）方法检测５１例散发性ＡＤ患者ＰＳ－１外显子８的３‘端内含子基因型,其中４０例晚发性ＡＤ患者,同时检测８７６例健康对照者ＰＳＤ－１基因型。结果发现散发性ＡＤ组１／１基因型和１等位基因频率分别为０．５１０和０．６９６,显著高于对照组的０．２９９和０．５２２;晚发性Ａ     

甲1型流感病毒新分离株HA基因的序列分析

目的 研究新分离的H1N1亚型流感病毒株的HA1基因序列。方法 甲型流感病毒通过鸡胚增殖后提取RNA、逆转录合成cDNA,经PCR扩增和产物纯化构建重组质粒,用双脱氧链终止法进行核苷酸序列测定;并进行基因特性分析。结果 新分离到的3株流感病毒株（H1N1）HA1区基因长度为981bp,编码327个氨基酸;与A/桂防/10/94和A/Bayern/07/95（H1N1）标准株比较其同源性分别为92.8%和91.3%,丢失了第130位氨基酸和304位糖基化位点;新分离的3株甲型流感病毒（H1地）标准株比较其同源性分别为92.8%和91.3%,丢失了第130位氨基酸和304位糖基化位点,新分离的3株甲型流感病毒株（H1N1）HA1区氨基酸同源性高达98%;A/桂防/10/94和A/Bayern/07/95(H1N1)毒株HN1氨基酸的同源性高达96%。结论 新分离到的3株H1N1毒株HA编码氨基酸不同于A/Baydrn/07/95(H1N1)和A/桂防/10/94（H1N1）标准株,它们可能为新的甲型流感病毒变异性。     

Cytochrome P450 1A1 and manganese superoxide dismutase genes polymorphisms in reactive arthritis

To investigate the role of cytochrome P450 1A1 (CYP1A1) and manganese superoxide dismutase (Mn SOD) genes polymorphisms in the susceptibility to reactive arthritis, the polymorphisms of CYP1A1 and Mn SOD genes were determined in 43 patients with reactive arthritis following Chlamydia trachomatis infection and 92 healthy controls by polymerase chain reaction (PCR)/restriction fragment length polymorphisms (RFLP) method.

The frequencies of CYP1A1 4887C/A and C/A+A/A were significantly higher in patients with reactive arthritis than in controls. Moreover, the increased frequency of CYP1A1 4887A in patients with reactive arthritis is independent of HLA-B27. The frequency of Mn SOD 1183T/T tended to be increased in patients with reactive arthritis. We also found that the frequency of CYP1A1 4887A was significantly increased in Mn SOD 1183T/T(+) reactive arthritis patients compared with 1183T/T(+) controls.

CYP1A1 4887A may be a risk factor for the development of reactive arthritis, especially in the presence of Mn SOD 1183T/T.     

Inefficient splicing of segment 7 and 8 mRNAs is an inherent property of influenza virus A/Brevig Mission/1918/1 (H1N1) that causes elevated expression of NS1 protein

Influenza A virus encodes two segments (7 and 8) that produce mRNAs that can be spliced. We have investigated if naturally occurring sequence polymorphisms in the influenza A virus family affects splicing of these viral mRNAs, as that could potentially alter the NS1/NS2- and/or M1/M2-protein ratios. We compared splicing efficiency of segment 7 and 8 mRNAs of A/Brevig Mission/1918/1 (H1N1) and A/Netherlands/178/95 (H3N2), as well as various H5N1 avian strains. Results revealed that both segment 7 and 8 mRNAs of A/Brevig Mission/1918/1 (H1N1) were inefficiently spliced compared to other influenza virus segment 7 and 8 mRNAs. This resulted in production of higher levels of functional NS1 protein, which could potentially contribute to the pathogenic properties of the A/Brevig Mission/1918/1 (H1N1). We also show that A/Brevig Mission/1918/1 (H1N1) segment 8 mRNAs responded differently to overexpression of SR proteins than A/Netherlands/178/95 (H3N2).     

Hemagglutinin antigenic determinants of influenza viruses H1N1 isolated 1953-1957 in the composition of the hemagglutinins of influenza viruses H1N1 isolated 1978-1979

Immunological analysis of antigenic determinants in the composition of hemagglutinins (HA) of H1N1 viruses isolated in the epidemic season of 1978-1979 showed their difference from A/USSR/090/77 virus HA. The A/Brazil/11/78 HA is characterized by 3 antigenic determinants two of which are identical with those of A/USSR/090/77 HA and one shows relationship with HA determinant of A/Omachi/1/53 virus, A/Fukushima/103/78 and A/Kagoshima/22/79 HAs are characterized by 1 determinant common with that of A/USSR/090/77 HA and 2 determinants identical with A/Omachi/1/53 virus HA. HAs of these viruses contain a determinant responsible for the antigenic relation of A/Omahi/1/53 HA with that of H1N1 viruses isolated in 1956-1957. The results suggest that the antigenic drift of HA of H1N1 viruses isolated in 1977-1979 occurs by the cycle characteristic of HA of H1N1 viruses isolated in 1950-1957. By the sensitivity to alpha-, beta-, and gamma-inhibitory of rabbit serum H1N1 viruses isolated in 1947-1957 and in 1977-1979 are quite similar with the exception of A/USSR/184/79 virus showing high sensitivity to gamma-inhibitors like H2N2 and H2N2 viruses.     

Polymorphism of DQA1 promoter in Italian population

We have investigated polymorphism in the 5′-URR of the DQA1 gene by PCR-SSO method in a group of 55 Italian healthy individuals olygotyped for DRB1, DQA1, DQB1 genes and in 20 10th IHWS cell lines as controls. We used primers and oligos (X and Y box) supplied by 12th IHWS and a DIG-11-ddUTP/AMPPD method. We have detected eight QAP variants (1.1,1.2,1.3,1.4,2.1,3.1,4.1,4.2) in our samples. As far as the association of DR/DQ haplotype and QAP sequences, we observed cases of one to one relationship (DQA1^*0201 and QAP2.1, DQA1^*0301 and QAP3.1, DQA1^*0401 and QAP4.2, DQA1^*0501 and QAP4.1); cases in which the same QAP allele was present in different DQA1-DRB1 haplotypes (QAP1.2 with DQA1^*0102 in DRB1^*15-DQB1^*0602 and DRB1^*16-DQB1^*0502 haplotypes or with DQA1^*0103 in the DRB1^*15-DQB1^*0601 haplotypes; QAP1.3 linked to DQA1^*0102, DQA1^*0103 or DQA1^*0104 in different haplotypes; QAP4.1 linked to DQA1^*0501 in DRB1^*11-DQB1^*0301, DRB1^*0301-DQB1^*0201, DRB1^*1303-DQB1^*0301 haplotypes or to DQA1^*0601 in DRB1^*0803-DQB1^*0301); cases where the same DQA1 allele is associated with different QAP sequences according to the DRB1 specificity (DQA1^*0102 allele with QAP1.2 or QAP1.4 in DRB1^*1302). Besides, we have observed that the QAP1.3, previously reported associated with DQA1^*0101-DRB1^*1401 haplotype, is really linked to DQA1^*0104-DRB1^*1401 haplotype. An intriguing data is that sometimes the same QAP is linked to different DQA1 alleles but to the same generic DRB1 allele: DRB1^*02 haplotype includes always the QAP1.2 variant but can bring different DQA1 alleles (^*0102 or ^*0103) and DRB1^*08 haplotype has always the QAP4.2 variant with different DQA1 alleles (^*0401 or ^*0601). The variability of linkage QAP-DQA1 can give further informations about HLA susceptibility in autoimmune diseases and in regulation of immune response in transplantation and oncology.