bluuebunny/crossref_metadata_2025 huggingface dataset:
Three novel MICB alleles
Authors: Fischer G. , Pérez‐Rodríguez M. , Argüello J.R. , Cox S.T. , McWhinnie A. , Travers P.J. , Madrigal J.A.
Published: March 2003

Acknowledgments:

This work is funded by the Anthony Nolan Bone Marrow Trust. M. P.‐R. is the recipient of a fellowship from Instituto Mexicano del Seguro Social.

The two members of the MHC class I chain‐related (MIC) gene family, MICA and MICB, have been shown by several investigators to be polymorphic. Most of the research effort so far has focussed on MICA, so less is known about the extent of polymorphism in the MICB gene. Here we report three novel MICB alleles, which had been detected in the course of an SSOP typing study on a large cohort of cell lines. Two of these alleles are formed by a non‐synonymous nucleotide variation. Our results confirm previous findings that most of the polymorphisms in the MICB gene, as in MICA, are coding and suggest that the extent of polymorphism in the two genes might be comparable.
Genetic polymorphisms in the cell growth regulated gene, SC1 telomeric of the HLA‐C gene and lack of association of psoriasis vulgaris
Authors: Teraoka Y. , Naruse T.K. , Oka A. , Matsuzawa Y. , Shiina T. , Iizuka M. , Iwashita K. , Ozawa A. , Inoko H.
Published: March 2003

Psoriasis vulgaris is associated with the HLA‐Cw6 and Cw7 antigens. We have previously narrowed down the critical region most likely to contain the psoriasis vulgaris gene to 111 kb spanning 89 kb to 200 kb telomeric of the HLA‐C locus by microsatellite mapping. This segment includes three known genes (POU5F1, SC1 and S) and four new expressed genes. Among them, SC1 (TCF19) is the cell growth regulated gene possibly with trans‐activator activity. Since psoriasis vulgaris is a common skin disorder characterized by hyperproliferation of epidermal cells, it is tempting to speculate that the SCI gene is one of the strong candidate genes responsible for the development of psoriasis vulgaris. Here, we investigated genetic polymorphisms in the SC1 gene by direct DNA sequencing and polymerase chain reaction‐single‐strand conformation polymorphism (PCR‐SSCP) techniques. Three single nucleotide polymorphisms in exon 2, two of which are accompanied by amino‐acid substitutions, were identified. Further, one 4‐bp deletion polymorphism was detected around the acceptor site of the lariat‐shaped structure necessary for RNA splicing in intron 2. No significant difference in the dimorphic or haplotypic distribution at these four polymorphic sites was observed between the patients with psoriasis vulgaris and healthy controls. This suggests that the susceptible gene for psoriasis vulgaris is not the SC1 gene itself, although a unique homozygous haplotype was identified in the patients( Note).
Seventeen more novel HLA‐A locus alleles
Authors: Ellis J. , Steiner N. , Kosman C. , Henson V. , Mitton W. , Koester R. , Ng J. , Hartzman R.J. , Hurley C.K.
Published: March 2003

This paper describes seventeen novel HLA‐A locus alleles: A*0106, *0235, *0236, *0237, *1105, *2302, *2303, *24032, *2422, *2424, *2503, *2613, *3007, *3203, *3204, *6809 and *6810. All alleles were identified due to unexpected probe reaction patterns during routine SSOP typing. Exons 2 and 3 of these alleles were subsequently characterized by DNA sequencing. The alleles represent a shuffling of sequence motifs, likely by interallelic conversion, expanding the diversity of the HLA system.
A novel HLA‐B*40 allele and novel exon 1 sequences of two B*40 alleles identified in potential marrow donors
Authors: Gourley I.S. , Wu J. , Bassinger S. , Montoya G.D. , Yee J. , Griffith B.B. , Kearns J. , McKeen M. , Birkos S. , Williams T.M. , Kamoun M.
Published: March 2003

Sequence‐specific oligonucleotide probe hybridization and sequence‐specific primer polymerase chain reaction (PCR) typing suggested the presence of variants of HLA‐B*40 in three individuals. Two were part of 3,500 potential marrow donors being screened for the National Marrow Donor Program, while the third was a clinical specimen. PCR products encompassing HLA‐B locus exons 1 through 3 were prepared and subcloned. In one individual, a native of the Pacific Islands, sequencing revealed a novel HLA‐B*40 allele (B*4023). In two other individuals, a previously unknown exon 1 sequence was determined for HLA‐B*4016 (ethnicity unknown) and B*4020 (Hispanic). These findings further illustrate the substantial genetic variation present at the HLA‐B locus within human populations ( Note).
HLA‐A*6817, identified in the Kolla Amerindians of North‐West Argentina possesses a novel nucleotide substitution
Authors: Ramon D. , Scott I. , Cox S.T. , Pesoa S. , Vullo C. , Little A.‐M. , Madrigal J.A.
Published: March 2003

Analysis of HLA polymorphism of the indigenous populations of Central and South America has identified many alleles not seen previously in other populations. We have described a novel allele, B*5113, previously in a Kolla Amerindian individual from North‐West Argentina. Here we present a second novel allele from this population: A*6817, which differs from its closest neighbour A*68012 by a single substitution at nucleotide 419. This substitution of adenosine 419 A*68012 for thymidine in A*6817 results in a novel amino acid change (aspartate to valine) at residue 116 ( Note).
Identification of a novel HLA‐Cw*05 allele, Cw*0503
Authors: Huang L.‐Q. , Boon T. , Van Pel A.
Published: March 2003

HLA‐Cw*05 is one of the least polymorphic subgroups of HLA‐C; so far only two alleles, namely Cw*0501 and Cw*0502, have been reported. We report here the identification of a third allele, Cw*0503, in a Caucasian individual. Cw*0503 is closely related to Cw*0501 with only six nucleotide substitutions clustering over a fragment of 48 nucleotides at the beginning of exon 4. All these six substitutions at the same positions have been found only in HLA‐B*44 alleles, suggesting that Cw*0503 is a result of recombination between Cw*0501 and one of B*44 alleles ( Note).
Major histocompatibility complex (MHC)‐ linked microsatellite markers in a founder population
Authors: Karell K. , Klinger N. , Holopainen P. , Levo A. , Partanen J.
Published: March 2003

The Finnish population is genetically relatively homogeneous and has a narrow gene pool as a result of founder effect followed by rapid population growth. We here demonstrate that microsatellite markers are highly informative tools for major histocompatibility complex (MHC) analysis in this population. First, no variation in 12 MHC‐linked microsatellites could be observed in certain CYP21‐deficient chromosomes, which as a result of founder effect most likely derived from common ancestors. Second, amongst 131 Finnish chromosomes, some, but not all, apparently HLA‐identical chromosomes also carried identical microsatellites, suggesting that these loci could be applied for identification of haplotypes which have a relatively recent shared origins. Finally, when the microsatellites were studied between ethnically more distant individuals (Finnish vs. non‐Finnish), who were matched for the HLA alleles, much more differences were observed. This showed that the similarity in microsatellites was population specific. The microsatellite typing can therefore be informative in fine mapping MHC‐linked susceptibility genes and can help in matching bone marrow transplants in isolated populations. Linkage disequilibrium was found to be much higher in the MHC than in another region (5q31) of similar size, indicating that there may be particular mechanisms keeping the MHC haplotypes conserved.
Allelic diversity of Mhc‐DRB alleles in rhesus macaques
Authors: Otting N. , De Groot N.G. , Noort M.C. , Doxiadis G.G.M. , Bontrop R.E.
Published: March 2003

The Biomedical Primate Research Centre (BPRC) rhesus macaque colony was started with a large number of wild‐caught animals originating mainly from the Indian subcontinent. The contemporary self‐sustaining colony comprises approximately 800 individuals. We screened a large section of the colony for Mamu‐DRB polymorphisms by applying the denaturing gradient gel electrophoresis (DGGE) technique. Based on disparate DGGE profiles, animals were selected for nucleotide sequence analysis. This approach allowed the detection of 25 unreported Mamu‐DRB alleles, bringing to 126 the total number of alleles documented in the literature. This communication demonstrates that rhesus macaques, like humans, display extensive allelic polymorphism at the DRB region. Phylogenetic analyses illustrate that humans and rhesus macaques share several Mhc‐DRB loci and lineages. Identical exon 2 sequences, however, which are shared between humans and rhesus macaques, were not observed. This indicates that most primate Mhc‐DRB alleles are of post‐speciation origin.
Genomic typing of minor histocompatibility antigen HA‐1 by reference strand mediated conformation analysis (RSCA)
Authors: Aróstegui J.I. , Gallardo D. , Rodríguez‐Luaces M. , Querol S. , Madrigal J.A. , García‐López J. , Grañena A.
Published: March 2003

Disparities in minor histocompatibility antigen (mHAg) HA‐1 are involved in the development of acute graft‐versus‐host disease (GvHD) in adult recipient after HLA‐identical sibling donor hematopoietic stem cell transplantation. The mHAg HA‐1 is an HLA‐A*0201‐restricted nonapeptide, which derives from the cleavage of a protein encoded at chromosome 19. The sequence analysis of HA‐1 cDNA identified two alleles, termed HA‐1H and HA‐1R, which differ in only two nucleotides at 3′ end of exon A, at positions 500 and 504. DNA‐based methods for HA‐1 typing were developed in 1998, using polymerase chain reaction with sequence‐specific primers (PCR‐SSP) and restriction fragment length polymorphism (PCR‐RFLP). Here, we report the usefulness of reference strand mediated conformation analysis (RSCA), which was developed for mutation detection and typing of polymorphic loci, to discriminate between the two HA‐1 alleles. We performed genomic typing of HA‐1 locus in 203 HLA‐A*0201‐positive samples using RSCA and we confirmed these results by PCR‐SSP. The results demonstrate the high reproducibility of this method and their strong correlation with the results obtained by PCR‐SSP (99%). Only two samples showed disparity between the RSCA typing and the PCR‐SSP. Direct sequencing of these samples confirmed that the correct allele assignment was that obtained by the RSCA typing. Furthermore, HA‐1‐ RSCA‐based typing provides additional information about the intronic structure of both alleles. With this approach, we describe the almost constant presence (99.2%) of a 5‐bp deletion at intronic position 214–218 associated to the HA‐1H allele, previously unidentified. We conclude that HA‐1 genomic typing by RSCA is easy to perform and that could be used as a routine typing method.
Reference strand mediated conformation analysis resolves HLA‐DRB1 typing ambiguities when matching for unrelated bone marrow transplantation
Authors: Corell A. , Pay A.L. , Little A.‐M. , Hoddinott M.A. , Argüello J.R. , Borton M. , Dunne C. , Ogilvie H. , O’Shea J. , Madrigal J.A. , Marsh S.G.E.
Published: March 2003

We show here the use of reference strand mediated conformation analysis (RSCA) to unambiguously resolve the HLA‐DRB1 typing of two individuals which were selected as potential unrelated donors for bone marrow transplantation (BMT). In the first case, both sequence‐specific primer (SSP) amplification and sequence‐specific oligonucleotide probing (SSO), routinely used in different tissue typing laboratories gave, for the two unrelated donors, the same ambiguous typing of HLA‐DRB1*04011+*0403 or DRB1*0407+*0413. In this case sequence‐based typing (SBT) was not the method of choice to resolve the situation, due to the sequence ambiguities of these two given combinations. RSCA of both samples, using homozygous typing cells (HTCs) for DRB1*04011, *0403 and *0407 as internal controls, gave the unambiguous result that both donors were HLA‐DRB1*04011+*0403. In the second case, a donor was typed as DRB1*1102+1103 by SSP, while SSO excluded the DRB1*1102 allele. The patient was unambiguously typed as DRB1*1101+1103 by both techniques. RSCA, using DNA from reference cell lines as internal controls, gave the unambiguous typing that the donor was DRB1*1103 homozygous.
Identification of two new alleles HLA‐DRB1*0312, DRB1*0432 and of a DRB3‐negative DRB1*1313‐positive haplotype
Authors: Anholts J.D.H. , Verduijn W. , Drabbels J. , Mulder A. , Doxiadis I.I.N. , Schreuder G.M.TH.
Published: March 2003

Two new HLA‐DRB1 alleles were identified in the course of routine class II molecular typing in Dutch Caucasoid. HLA‐DRB1*0312 is similar to *03011 except for codon 57 (GAT→AGC). DRB1*0432 is similar to *0413 but with a mutation at position 215, changing codon 72 (CGG→CAG; Arg→Gln). This sequence has never before been identified at this position. A DRB3‐negative DRB1*1313 haplotype was identified in an individual from Indonesia. Monoclonal antibodies against DR52 were non‐reactive with lymphocytes of this individual. The DRB1*1313‐DRB3‐negative haplotype probably represents a recombination of DRB1*13 and *08 haplotypes where the sequences telomeric of HV1 are derived from the DRB3‐negative DRB1*0803 haplotype ( Note).
Lectin purified human class I MHC‐derived peptides: evidence for presentation of glycopeptides in vivo
Authors: Kastrup I.B. , Stevanovic S. , Arsequell G. , Valencia G. , Zeuthen J. , Rammensee H.‐G. , Elliott T. , Haurum J.S.
Published: March 2003

Previously, using synthetic glycopeptides carrying a natural cytosolic type of monosaccharide O‐β‐linked N‐acetylglucosamine (GlcNAc) glycosylation of serine residues, we have shown that glycopeptides act as suitable substrates for TAP‐mediated transport into the endoplasmic reticulum (ER), and that they bind efficiently to class I major histocompatibility complex (MHC) molecules and can elicit glycopeptide‐specific cytotoxic T‐lymphocyte (CTL) responses in mice. Recently, we have reported that peptides presented by human class I MHC molecules in vivo encompass a small but significant amount of peptides which seem to be carrying O‐β‐linked monosaccharide GlcNAc. In the present report we provide further evidence that glycosylated peptides are indeed presented by class I MHC molecules in vivo. Thus, peptides derived from HLA‐A*0201 were purified by wheat germ agglutinin (WGA) lectin affinity chromatography as previously described. Subsequently, the peptides contained in the WGA‐eluate were subjected to sequence analysis by Edman degradation. It was found that the peptides derived from HLA‐A*0201 which had been retained by the O‐GlcNAc‐binding lectin WGA did indeed carry a HLA‐A*0201 binding motif. Furthermore, using an enzymatic labeling procedure we present evidence that the HLA‐A*0201‐derived peptides which bind to the WGA lectin are glycosylated with terminal GlcNAc residues. Together, these data provide further evidence for the natural presentation by human class I MHC of glycopeptides carrying terminal O‐GlcNAc residues in vivo.
Polymorphism of the ABO blood group genes in Han, Kazak and Uygur populations in the Silk Route of northwestern China
Authors: Iwasaki M. , Kobayashi K. , Suzuki H. , Anan K. , Ohno S. , Geng Z. , Li G. , Inoko H.
Published: March 2003

Genetic polymorphism in the ABO blood group gene of Han, Kazak and Uygur populations inhabiting the most northwestern part of China was investigated using polymerase chain reaction‐based techniques. The present study enrolled 43 healthy unrelated Han, 37 Kazak and 59 Uygur volunteers. The allele in A1 blood group is distinguished A0101 and A0102 in difference of nucleotide position 467. The A0101 allele is more frequent in Caucasian and the A0102 allele is characteristic in Mongoloid. It must be notable that A0201 in the A2 group (with a single base deletion at nucleotides 1059 to 1061) which was characteristic of Caucasian was observed in Kazak and Uygur populations but not in Han. Further, O0201 (with no nucleotide deletion at 261 and three nucleotide differences), which is frequent in different races including Caucasian except for Mongoloid, was detected also in Kazak and Uygur populations. The frequencies of B0101 in Kazak, Uygur and Han were comparable to those of other Asian populations but higher than those of Caucasian populations. Collectively, these results reveal that the allele frequencies of Kazak and Uygur at the ABO blood group locus are an intermediate between those of Mongoloid and Caucasian, suggesting the admixed feature of Kazak and Uygur with Mongoloid and Caucasian.
Characterization of HLA‐B*3921 and confirmation of HLA‐B*4415, two variant HLA‐B alleles identified in the Omani population
Authors: Williams F. , Curran M.D. , Leheny W.A. , Daar A.S. , Middleton D.
Published: March 2003

We describe a variant HLA‐B*39 allele present in two individuals from Oman, which has been officially named HLA‐B*3921. In addition we confirm the existence of HLA‐B*4415, an allele closely related to HLA‐B*4501 differing only at the Bw4/Bw6 epitope ( Note).
Peptide specificity of the Amerindian B*3905 allotype: molecular insight into selection mechanisms driving HLA class I evolution in indigenous populations of the Americas
Authors: Yagüe J. , Ramos M. , Ogueta S. , Vázquez J. , López de Castro J.A.
Published: March 2003

HLA‐B*3905 is apparently restricted to Amerindian populations and presents a wide geographical distribution, from Mexico to Argentina. It differs from B*3901, one of the founder HLA class I alleles of Central and South Amerindians, by a single nucleotide substitution leading to an Asp74Tyr change in the gene product. The peptide specificity of the B*3905 protein was characterized by pool sequence analysis of B*3905‐bound peptides and by sequencing of a set of individual ligands, using electrospray ion trap mass spectrometry. The results indicate a double effect of the B*3905 mutation. First, pocket B specificity was shifted towards an increased preference for His at peptide position 2, which is the main anchor for B39‐bound peptides, relative to B*3901. Second, at peptide position 3 acidic residues were favored, and aromatic residues disfavored, relative to B*3901. These features approach the peptide specificity of B*3905 to B*3801 and B*1509, allotypes absent from Central and South Amerindians. Together with B*3909, B*3905 is the second HLA‐B39 subtype whose polymorphism results in a shift of peptide specificity towards that of HLA‐B allotypes absent from these populations. This suggests that HLA‐B39 evolution in Central and South America may be an antigen‐driven adaptive response, leading to generate antigen‐presenting properties absent from the HLA class I repertoire of the ancestral population.
Sequence analysis of a new HLA‐DR11 allele in a Caucasian Italian family: DRB1*11272
Authors: Canossi A. , Piancatelli D. , Papola F. , Di Rocco M. , Del Beato T. , Liberatore G. , Casciani C.U. , Adorno D.
Published: March 2003

Acknowledgments:

This research was supported in part by funding from the C.N.R., P.F. “Biotecnologie”. We wish to acknowledge and thank Dr. Maria Teresa Vicentini, Dr. Lorena Pompucci and Dr. Anna Aureli for their support with HLA typing.

This communication describes a new DRB1*11 allele identified in a familial group of Caucasian individuals from central Italy. The new sequence has been officially named DRB1*11272 by the World Health Organization (WHO) Nomenclature Committee (February 2000). It encodes an HLA antigen serologically recognized as DR11. The sequence variation of this new allele was localized to codon 77 of exon 2. It differs at position 230 from DRB1*11011 allele (A replacing C), and at position 231 from DRB1*1127 (C replacing T). At the amino acid level, the DRB1*11272 mutation is silent with respect to the DRB*1127 phenotype, coding for a synonymous asparagine.
Functional topography of discrete domains of human CD38
Authors: Ausiello C.M. , Urbani F. , Lande R. , La Sala A. , Di Carlo B. , Baj G. , Surico N. , Hilgers J. , Deaglio S. , Funaro A. , Malavasi F.
Published: March 2003

In addition to its catalytic activities as ecto‐NAD+ glycohydrolase (NADase), CD38 displays the ability to transduce signals of biological relevance. Indeed, ligation of CD38 on peripheral blood mononuclear cells (PBMC) by agonistic monoclonal antibodies (mAbs) is followed by the transcription and secretion of a vast array of regulatory cytokines. The present work addresses the issue of whether the signals leading to calcium (Ca2+) mobilization, lymphocyte proliferation and release of cytokines is dependent on the epitopes recognized by the individual mAbs. Competition binding analysis identifies two families of mAbs, namely IB4, IB6 and AT2 on one side and OKT10, SUN‐4B7 and AT1 on the other. Each mAb family binds epitopes that are completely or partially common. However, the functional activities of the CD38 molecule can not be simply attributed to the epitopes engaged: for instance, IB4 and OKT10 mAbs, which bind different epitopes, perform as agonistic mAbs in inducing PBMC proliferation and interferon (IFN)‐γ secretion. SUN‐4B7 yields intermediate effects, whereas IB6, AT1 and AT2 mAbs are totally ineffective. The effects mediated by IB4 and OKT10 mAbs are apparent in 80% of the healthy individuals studied, whereas the effects of SUN‐4B7 mAb operate only in 25% of the donors. Interleukin (IL)‐6 secretion was observed in all individuals analyzed, irrespective of the epitopes triggered and of mAbs used to ligate the CD38 molecule. In addition, IB4 is the only mAb able to induce significant intracellular Ca2+ fluxes.
HLA‐DRB1*03, DRB1*11 or DRB1*12 and their respective DRB3 specificities in clinical variants of sarcoidosis
Authors: Bogunia‐Kubik K. , Tomeczko J. , Suchnicki K. , Lange A.
Published: March 2003

Determination of DRB1 and DRB3 specificities in sarcoidosis patients identified that the presence of DRB1*03 and the absence of DRB1*11 and/or DRB1*12 favors a course of disease that is associated positively with Löfgren’s syndrome (DRB1*03) and negatively with stage I disease (DRB1*11 and/or 12). In common with normal controls, DRB1*03 was associated with DRB3*0101 and DRB1*11/12 with DRB3*0201/2. An analysis of DRB1 and DRB3 associations in variants of sarcoidosis revealed that DRB1*03 and DRB3*0101 were associated with Löfgren’s syndrome in a combined association fashion. Conversely, a lack of DRB1*11 and/or DRB1*12 but not DRB3*0201/2 favored the clinical course of sarcoidosis.
New alleles in the B44 family including B*44022, B*44032, B*4411, B*4420, B*4421, B*4424, and B*8301
Authors: Steiner N.K. , Gans C.P. , Kosman C. , Bradshaw D. , Koester R. , Menchaca E.M. , Mitton W. , Ng J. , Hartzman R.J. , Hurley C.K.
Published: March 2003

Abstract: Seven new HLA‐B locus alleles have been described. B*44022 and B*44032 are silent substitutions altering known alleles. B*4411 carries a unique Bw4‐like epitope. B*4420, B*4421, and B*4424 carry new combinations of motifs previously observed in other alleles. B*8301 appears to be the result of the replacement of exon 2 from B*4402 with exon 2 from B*5603.
Identification of two new HLA‐B22 variants, HLA‐B*5509 and B*5606
Authors: Voorter C.E.M. , Hepkema B.G. , Mulkers E.M.T. , Lems S.P.M. , Van Den Berg‐Loonen E.M.
Published: March 2003

Abstract: In our recent study using high‐resolution HLA‐B locus typing by sequence‐based typing (SBT) we identified 9 new alleles in a total of 355 unrelated individuals (4). Three of them concerned an allele belonging to the B22 group. One of them, B*5607, showed the unusual presence of a Bw4 sequence motif, as described previously (5). In this report the other two B22 variants are described; one belonging to the B55 specificity and named B*5509; the other one being a B*56 allele and assigned B*5606, which brings the total number of alleles belonging to the B22 group to 18.
Nucleotide sequence of exons 2 and 3 of the novel HLA‐Cw*12032 allele
Authors: Velickovic Z.M. , Carter J.M.
Published: March 2003

Abstract: A novel HLA‐C nucleotide sequence was discovered in an individual of Pacific Island, Tongan ethnicity using sequencing based typing. The sequence was given an official allele designation HLA‐Cw*12032 by the WHO Nomenclature Committee for Factors of the HLA System. The novel sequence has 2 bp substitutions compared to the HLA‐Cw*1203 at codon 134 “T” to “C” and codon 135 “C” to “G”. However, when converted to amino acid sequences HLA‐Cw*1203 and HLA‐Cw*12032 are identical and would not have direct clinical implications.
Identification of a novel HLA‐B*2722 allele from a Filipino cell
Authors: Witter K. , Lau M. , Zahn R. , Scholz S. , Albert E.D.
Published: March 2003

Abstract: We present the novel HLA‐allele B*2722 amplified and sequenced from a Filipino individual. This DNA was included several times in the International Cell Exchange, UCLA, and typed at the time as B*27 or B*2706 by the majority of the participating laboratories. The second HLA‐B allele of this person is B*3802. B*2704/06 or B*2704/06/10 were suggested by approximately one‐third of the laboratories. As we were investigating the intron 3 sequences of B*27 alleles, we also sequenced exon 4 of this Filipino DNA and found a single nucleotide exchange in exon 4 (pos. 704) which was not in concordance with the previous B*2706 typing. Our sequencing results showed that the coding sequence of B*2722 is identical to B*2706 in exon 1, 2 and 3. In exon 4 the B*2722 sequence is identical to B*2704. HLA‐B*2704 differs from B*2706 in one base at position 704, a G in B*2704 and a C in B*2706, respectively.
ANALYSIS OF THE RESULTS OF PAIRED CADAVERIC KIDNEYS WITH SIMILAR DONOR‐RECIPIENT HLA MATCHINGS
Authors: Yaǧmurdur MC , Emiroğlu R , Basaran O , Karakayalı H , Tezel E , Haberal M
Published: March 2003

In this study, we aimed to clarify the factors that may effect long‐term graft and patient survivals. We retrospectively evaluated the outcome of 112 renal transplant recipients who shared paired cadaveric kidneys between 1985 and July 2001.

There were two study groups, each with 56 patients, according to the timing of the operations. The operations were performed by the same transplant team, so Group 1 represents the recipients of the first kidney and Group 2, the second. The same low‐dose triple drug immunosuppression protocol with prednisolone, azathioprine and cyclosporine A was used in both groups. Cyclosporine A was converted to tacrolimus in seven patients in Group 1 and in five patients in Group 2. Demographic properties, number of acute‐rejection episodes, serum creatinine levels, creatinine clearance values, graft and patient survival rates, postoperative complications, recipient body weights, cold ischemia times, and de novo malignancies were studied. To analyze our results the Chi‐square and Wilcoxon Signed Ranks tests were used, differences, with P<0.05 were accepted as significant.

Regarding the HLA matchings, the groups were similar. Mean cold ischemia times were 22.1±8.7 and 23.3±9.1 h in Groups 1 and 2, respectively (P>0.05). Acute‐rejection rates within the first month following the transplantation were also similar in both groups. Serum creatinine levels and creatinine clearance values at 30 days post‐transplantation showed no significant difference between the groups (P>0.05). One, 2 and 5 years' graft survival rates (Group 1 vs. Group 2) were 97% vs. 92%, 74% vs. 77% and 27% vs. 24%, respectively. No statistically significant difference was observed on returning to the hemodialysis rates (10/56 vs. 13/56). In Group 2, the rate of proteinuria (over 200 mg/day) throughout the follow‐up period was significantly higher than that in Group 1 (Chi‐square, P=0.03). Three patients underwent re‐transplantation: one in Group 1 and two in Group 2. Three post‐transplant malignancies were detected: one in Group 1 and two in Group 2. An interesting observation was the post‐transplantation T‐cell lymphoma diagnosis in both recipients of the same donor. Also, when compared, the patients were not significantly statistically different according to their body weights, rejection rates and graft survival rates.

In conclusion, the short‐ and long‐term outcomes of consecutive‐paired cadaveric kidney transplantations are comparable with similar cold ischemia times, HLA matching and immunosuppressive protocols.
The cell biology of MHC class I antigen presentation
Authors: Williams A. , Peh C. A. , Elliott T.
Published: March 2003

Abstract: MHC class I antigen presentation refers to the co‐ordinated activities of many intracellular pathways that promote the cell surface appearance of MHC class I/β2m heterodimers loaded with a spectrum of self or foreign peptides. These MHC class I peptide complexes form ligands for CD8 positive T cells and NK cells. MHC class I heterodimers are loaded within the endoplasmic reticulum (ER) with peptides derived from intracellular proteins. Alternatively, MHC class I molecules may be loaded with peptides derived from extracellular proteins in a process called MHC class I cross presentation. This pathway is less well defined but can overlap those pathways operating in classical MHC class I presentation and has recently been reviewed elsewhere (1). This review will address the current concepts regarding the intracellular assembly of MHC class I molecules with their peptide cargo within the ER and their subsequent progress to the cell surface.
Extensive interbreed, but minimal intrabreed, variation of DLA class II alleles and haplotypes in dogs
Authors: Kennedy L. J. , Barnes A. , Happ G. M. , Quinnell R. J. , Bennett D. , Angles J. M. , Day M. J. , Carmichael N. , Innes J. F. , Isherwood D. , Carter S. D. , Thomson W. , Ollier W. E. R.
Published: March 2003

Abstract: The DLA class II genes in the dog major histocompatibility complex are highly polymorphic. To date, 52 DLA‐DRB1, 16 DLA‐DQA1 and 41 DLA‐DQB1 allelic sequences have been assigned. The aim of this study was to examine the intrabreed and interbreed variation of DLA allele and haplotype frequencies in dogs, and to ascertain whether conserved DLA class II haplotypes occur within and between different breeds. One thousand and 25 DNA samples from over 80 different breeds were DLA class II genotyped, the number of dogs per breed ranging from 1 to 61. DNA sequence based typing and sequence specific oligonucleotide probing were used to characterize dogs for their DLA‐DRB1, DQA1 and DQB1 alleles. The high frequency of DLA class II homozygous animals (35%), allowed the assignment of many haplotypes despite the absence of family data. Four new DLA alleles were identified during the course of this study. Analysis of the data revealed considerable interbreed variation, not only in allele frequency, but also in the numbers of alleles found per breed. There was also considerable variation in the number of breeds in which particular alleles were found. These interbreed variations were found in all three DLA class II loci tested, and also applied to the three‐locus haplotypes identified. Within this data set, 58 different DLA‐DRB1/DQA1/DQB1 three‐locus haplotypes were identified, which were all found in at least two different animals. Some of the haplotypes appeared to be characteristic of certain breeds. The high interbreed, and relatively low intrabreed, variation of MHC alleles and haplotypes found in this study could provide an explanation for reports of interbreed variation of immune responses to vaccines, viruses and other infections.
Association of NRAMP1 promoter gene polymorphism with the susceptibility and radiological severity of rheumatoid arthritis
Authors: Rodríguez M.R. , González‐Escribano M.F. , Aguilar F. , Valenzuela A. , García A. , Núñez‐Roldán A.
Published: March 2003

Abstract: The natural resistant‐associated macrophage protein 1 (NRAMP1) has been proposed as a candidate gene for the susceptibility to autoimmune diseases. In this study, the possible role of the functional polymorphism located at the promoter region of NRAMP1 gene in the susceptibility and clinical outcome of rheumatoid arthritis (RA) was investigated. A total of 141 Spanish RA patients and 194 controls previously typed for HLA‐DRB1* were genotyped for the NRAMP1 polymorphism. No significant differences in the distribution of frequencies among RA patients and controls were observed. Nevertheless, when patients and controls were stratified according to their HLA shared epitope (SE) status, an increase of 2/2 genotype among SE‐negative (SE‐) patients with respect to SE‐ controls was observed (23% vs 7%, OR = 3.74, 95% CI 1.31–10.72). In addition, the possible role of this polymorphism in the clinical course of RA was investigated in a subgroup of 82 patients who were prospectively followed during a mean of 9 years. After follow‐up, an increase of patients with the homozygous 2/2 genotype was detected among those with severe small joint radiological involvement: 73% of patients 2/2 had a severe form in contrast to 37% of patients with the genotype 2/3 and 30% of patients bearing 3/3 OR = 5.45, 95% CI 1.14–34.24). In conclusion, NRAMP1 gene promoter polymorphism could influence the radiological severity of rheumatoid arthritis and disease susceptibility, particularly in individuals lacking HLA‐linked risk factors.
HLA‐B*3531, a hybrid of B35 and B61, implications for diagnostic approaches to alleles with complex ancestral compositions
Authors: Elsner H.‐A. , Himmel A. , Steitz M. , Hammer P. , Schmitz G. , Ballas M. , Blasczyk R.
Published: March 2003

Abstract: The serological characterization of allelic variants that have been generated by large‐scale interallelic recombination events indicates which residues may be involved in the formation of epitopes crucial for serological recognition. The allelic product of HLA‐B*3531 is composed of B35 in its α1 domain and of B61(40) in its α2 domain. Both specificities are only weakly detectable with available sera. Allelic products with ‘mixed’ serology also represent a challenge to DNA‐based HLA typing methods, as only the sequence motif of one ancestral allele may be recognized. In this case the hidden specificity would not be considered in the matching process and might not be recognized as an antigen ‘unacceptable’ to the recipient.
HLA‐G polymorphisms in couples with recurrent spontaneous abortions
Authors: Hviid T.V. , Hylenius S. , Hoegh A.M. , Kruse C. , Christiansen O.B.
Published: March 2003

Abstract: The etiology of a fraction of recurrent spontaneous abortions (RSA) may involve immunological mechanisms. Aberrant profiles of Th1 and Th2 cytokines have been observed which are not present in uncomplicated pregnancies. Studies of classical HLA class I and II antigens in relation to RSA have not been conclusive. Furthermore, these antigens are not expressed in the placenta with the exception of HLA‐C. However, HLA‐G is expressed on especially invasive cytotrophoblasts and exists in both membrane and soluble forms. HLA‐G may be involved in materno‐fetal tolerance. Therefore, 61 RSA couples (with three or more spontaneous abortions) and 47 fertile control couples were HLA‐G genotyped by direct DNA sequencing and analyzed for specific polymorphisms. No statistically significant differences were observed in the distribution of HLA‐G alleles between controls and RSA couples, however, 15% of the RSA women carried the HLA‐G*0106 allele compared to 2% of the control women. The 14 bp deletion polymorphism in exon 8 was investigated separately. There were a greater number of heterozygotes for the 14 bp polymorphism in the group of fertile control women than expected, according to Hardy–Weinberg equilibrium. Furthermore, the HLA‐G alleles without the 14 bp sequence were prominent in the RSA males in contrast to the RSA women in whom alleles including the 14 bp sequence were frequently observed, especially as homozygotes. These results are discussed in relation to two hypotheses concerning HLA‐G and RSA. A hypothesis of HLA‐G histo‐incompatibility between fetus/placenta and the mother was not supported by the data. Another hypothesis concerned certain HLA‐G alleles associated with an altered expression profile of HLA‐G isoforms or reduced expression of certain HLA‐G isoforms.
Antibacterial activity of multiple antigen peptides homologous to a loop region in human lactoferrin
Authors: Azuma M. , Del Carpio, C.A. , Kojima T. , Yokoyama, I. , Tajiri, H. , Yoshikawa K. , Saga, S.
Published: March 2003

Abstract: An 11‐residue peptide (FQWQRNMRKVR) homologous to just over half the loop region of human lactoferricin is thought to be responsible for antimicrobial properties of human lactoferricin. Multiple antigen peptides (MAP) of the 11‐residue peptide exerted significant antibacterial effects against a broad spectrum of bacteria including MRSA. More than eight branching was favourable for increasing its antibacterial activity. Our report shows a novel possibility for MAP to increase the activity of antibiotic peptides other than simply to stimulate antibody production, as reported so far.
Effects of temporins on molecular dynamics and membrane permeabilization in lipid vesicles
Authors: Rinaldi A.C. , Di Giulio A. , Liberi M. , Gualtieri G. , Oratore A. , Bozzi A. , Schininà M.E. , Simmaco M.
Published: March 2003

Abstract: Temporins are a novel family of small (10–13 residues) cationic antimicrobial peptides recently isolated from the skin of the European red frog Rana temporaria. Although recently acquired evidence shows that temporins have the potential to kill bacteria by permeabilizing the cytoplasmic membrane, the molecular mechanisms of membrane selectivity and permeabilization are largely unknown. In this study, it was found that temporins cause the release of fluorescent markers entrapped in phosphatidylcholine liposomes in a manner that depends significantly on the size of the solute. Temporins were also shown to lack a detergent‐like effect on lipid vesicles, indicating that marker leakage caused by these peptides is not due to total membrane disruption but to perturbation of bilayer organization on a local scale. Binding of temporins to liposomes did lead to a small increase in lipid hydrocarbon chain mobility, as revealed by EPR spectroscopy of nitroxide‐labeled fatty acids incorporated in the bilayer. Reference experiments were conducted using the bee venom peptide melittin, whose properties and behavior in natural and model membrane systems are well known. Our findings for temporins are discussed in relation to the models proposed to date to account for the action of antimicrobial peptides on membranes.
Fast, efficient and selective deprotection of the tert‐butoxycarbonyl (Boc) group using HCl/dioxane (4 m)
Authors: Han G. , Tamaki M. , Hruby V.J.
Published: March 2003

Abstract: Fast, efficient and selective deprotection of the tert‐butoxycarbonyl (Boc) group of various amino acids and peptides was achieved by using hydrogen chloride (4 m) in anhydrous dioxane solution for 30 min at room temperature. In the cases studied in our laboratory, this protocol provided superior selectivity to deprotect Nα‐Boc groups in the presence of tert‐butyl esters and tert‐butyl ethers, including thio‐tert‐butyl ethers, but not phenolic tert‐butyl ethers.
Measurement of the affinity of melittin for zwitterionic and anionic membranes using immobilized lipid biosensors
Authors: Lee T.‐H. , Mozsolits H. , Aguilar M.‐I.
Published: March 2003

Abstract: The binding of melittin to zwitterionic dimyristyphosphatidylcholine (DMPC) and anionic dimyristylphosphatidylglycerol (DMPG) was analysed using two different immobilized model membrane systems. The first system used surface plasmon resonance (SPR), which monitors the real‐time binding of peptides to an immobilized hybrid bilayer. SPR experiments reflected a stronger binding of melittin for DMPG than for DMPC, while kinetic analysis suggested the existence of at least two distinct binding steps. The second lipid biosensor system involved an immobilized phospholipid monolayer covalently attached to a microporous silica surface. The binding of melittin to the immobilized monolayer was then monitored using dynamic elution chromatography with varied methanol concentrations to analyse the binding of melittin to DMPC and DMPG. The nonlinear binding behaviour observed for melittin with the phosphatidylcholine (PC) and phosphatidylglycerol (PG) monolayers compared with the linear retention plots and Gaussian peak shapes observed for the control molecule demonstrated that melittin undergoes significant conformational and orientational changes upon binding to the immobilized PC and PG ligands. The dependence of log k′ on per cent methanol also demonstrated a bimodal interaction whereby hydrophobic forces predominated at higher temperatures and methanol concentrations, while other forces, presumably electrostatic in nature, also made a contribution to the affinity of the peptides for the lipid monolayer, particularly at lower temperatures. The complementary use of these two lipid biosensors thus allows the role of hydrophobic and electrostatic forces in peptide–membrane interactions to be studied.
Studies on the binding of Treponema pectinovorum to HEp‐2 epithelial cells
Authors: Walker S. G. , Ebersole J. L. , Holt S. C.
Published: March 2003

We developed a radioassay to assess the adherence of the oral treponemes Treponema denticola and Treponema pectinovorum to live HEp‐2 epithelial cells. T. pectinovorum bound firmly to the epithelial cell monolayer in a concentration‐dependent manner. The results indicated that a subpopulation of T. pectinovorum appeared to bind and that the binding could be influenced by environmental factors. Increasing concentrations of fetal bovine serum inhibited binding, whereas T. pectinovorum membrane vesicles and co‐incubation with T. denticola ATCC 35404 increased the number of cells bound to the monolayer. Treatment of T. pectinovorum with periodic acid, but not trypsin or proteinase K, decreased the binding suggesting that a cell surface carbohydrate, such as the O‐antigenic component of the lipopolysaccharide, mediates attachment of the bacteria to the epithelial cells. Co‐infection of the HEp‐2 cells with both T. denticola and T. pectinovorum did not interfere with each other in attachment to the epithelial cell suggesting that they do not compete for the same cellular receptor on the host cell surface. This study demonstrates that T. pectinovorum is capable, in vitro, of forming a tight association with host cells and that this binding could represent an initial step in the pathogenesis of T. pectinovorum.
Serum antibody response to oral infection precedes but does not prevent Porphyromonas gingivalis–induced alveolar bone loss in mice
Authors: Baker P. J. , Carter S. , Dixon M. , Evans R. T. , Roopenian D. C.
Published: March 2003

The purpose of this study was to determine whether humoral immunity prevents bacterially induced alveolar bone loss. BALB/cByJ mice were orally infected with the human periodontopathic bacterium Porphyromonas gingivalis, and were compared with sham‐infected mice. Specific serum antibody titers to P. gingivalis were measured by enzyme‐linked immunosorbent assay. Alveolar bone levels were measured as the distance from the cementoenamel junction to the alveolar bone crest and bone loss was defined as a change in bone levels over time or between infected and sham‐infected animals. The specific humoral response was predominantly of the IgG isotype, although low levels of specific serum IgA were also present. Antibody titers in the infected animals were significantly different from those in the sham‐infected animals by 18 days and remained at maximal levels at 47 days. Bone loss became significant by 26 days and continued to progress at 47 days. Thus the serum antibody response to oral infection with P. gingivalis preceded detectable bone loss and remained elevated while bone loss increased. The presence of specific antibody did not prevent the onset or progression of bone loss.
Role of transcriptional factor activation protein‐1 in endogenous expression of the interleukin‐1β gene involved in Porphyromonas gingivalis fimbria–stimulated bone resorption in the mouse calvarial system
Authors: Naganuma K. , Amano S. , Takeda H. , Kitano S. , Hanazawa S.
Published: March 2003

We previously suggested that endogenous interleukin‐1β (IL‐1β) produced by Porphyromonas gingivalis fimbriae stimulation in calvarial bone cell cultures plays a role in bone resorption as a major cytokine. Therefore, in the present study, we initiated experiments to clarify the stimulatory mechanism of IL‐1β gene expression in fimbria‐stimulated bone resorption. Fimbria‐stimulated bone resorption was dramatically inhibited by curcumin, a potent inhibitor of activation protein‐1 (AP‐1). In fact, the fimbriae induced markedly both the expression of c‐fos and c‐jun genes and their protein production in the calvarial cells. In addition, a mixture of antisense oligonucleotides against c‐fos and c‐jun significantly inhibited not only the fimbria‐induced expression of the IL‐1β gene but also the fimbria‐induced bone resorption. Therefore, the present study demonstrates that transcriptional factor AP‐1 plays a functional role in P. gingivalis fimbria–stimulated bone resorption via endogenous IL‐1β in the mouse calvarial system.
Identification and characterization of human immunoglobulin G Fc receptors of Fusobacterium nucleatum
Authors: Guo M. , Han Y. W. , Sharma A. , De Nardin E.
Published: March 2003

Several human pathogens express components which can bind to the Fc portion of immunoglobulins. This study was undertaken to characterize the human immunoglobulin G (IgG) Fc‐binding activity of Fusobacterium nucleatum, a suspected pathogen involved in periodontal diseases. Fc‐binding activity was detected using whole‐cell, cell envelope and outer membrane fractions, and it was found to be associated with polypeptides of 40 kDa and 42 kDa, respectively. Amino terminal sequencing of these components revealed them to be homologous to the bacterial porin encoded by fomA gene. Further sequencing of internal peptide fragments obtained by CNBr cleavage suggested that these two proteins are probably isoforms. In summary, we show that a porin‐like protein on the surface of F. nucleatum can bind the Fc fragment of the human immunoglobulin G, and this protein may act as a virulence factor to facilitate this bacterium in evading host immune surveillance system.
Antigenic cross‐reactivity among Porphyromonas gingivalis serotypes
Authors: Fan Q. , Sims T. J. , Nakagawa T. , Page R. C.
Published: March 2003

The goal of our research program is to develop a Porphyromonas gingivalis vaccine. Vaccine development requires identification of antigenic components shared by the many clonal types of P. gingivalis. The purpose of the present study was to evaluate the extent and nature of antigenic cross‐reactivity among serotypes of P. gingivalis and to identify shared antigenic components. Strains selected to represent serotypes A–D were 33277, A7A1‐28 W50 and 381, respectively. Using intact cells, antibodies were raised in rabbits. Titers were assessed by enzyme‐linked immunosorbent assay (ELISA) using intact cells as antigen, Western blots were prepared and biologic activity was measured as opsonization (chemiluminescence expressed as mV) and enhancement of phagocytosis and killing by polymorphonuclear leukocytes. Extensive cross‐reactivity that varied greatly among serotypes was observed by ELISA. The Western blots showed an even greater extent of cross‐reactivity, with shared protein components at approximately 140, 130, 37, 32 and 28 kDa and a shared variable molecular mass smear considered to be lipopolysaccharide and other carbohydrate. Additional protein components at 110, 85, 35 and 20 kDa appeared to be shared by some but not all serotypes. In the functional assays, strains 33277 and 381 were equally well opsonized by anti‐33277 and anti‐381 (500–650 mV) but opsonized to a much lesser extent by anti‐A7A1‐28 and anti‐W50 (roughly 125 mV and 350 mV respectively). A7A1‐28 and W50 were opsonized by all four immune sera almost equally but to a much lower extent (roughly 400 mV and 250 mV respectively). Enhancement of phagocytosis and killing in the presence of active complement mirrored opsonization with the exception that 381 was reasonably well opsonized by anti‐A7A1‐28 (400 mV) and anti‐W50 (350 mV), but poorly killed. The protein components at 140, 130, 37 and 28 kDa shared by all of the four serotypes appear to have potential as vaccine candidate antigens.