Hi,
We typed (our) HeLa as HLA-A*6802, -B*1503 or -B*1537, and -Cw*1604
using RT-PCR [Johnson and Mook-Kanamori, JBC 275 (22) 16643-9 (2000)].
These were the best matches from single sequencing runs of PCR amplified
cDNAs, not rigorously checked. HeLa have been serologically typed as
HLA-A3, -A28, -Bw15, and Bw35 [Espmark et al Tissue Antigens 11:279-86
(1978)] and genetically typed as HLA-A68 (a subtype of A28), -B75 (a
subtype of B15) and NEGATIVE for B35 [Benham, Gromme and Neefjes,
J.Immunol. 161:83-89 (1998)].
Obviously, these findings disagree. You might consider HLA class I
typing your own HeLa with the RT-PCR procedure that we described
[Johnson, Biedermann and Mook-Kanamori, J. Immunol. Meth. 233:119-129
(2000)]. It's very easy, quick, and requires only 4 primers.
I hope this helps.
Dave
John Ladasky wrote:
>iayork at panix.com (Ian A. York) wrote in message news:<adm38p$sfb$1 at reader1.panix.com>...
>>>Does anyone know the MHC class I alleles expressed by HeLa cells? I've
>>found two HLA-A alleles associated with HeLa in Genbank, but I haven't
>>found any of the HLA-B alleles. Someone must have looked, somewhere, wome
>>time.
>>>>Thanks.
>>>>Ian
>>> Hi, Ian!
>> We seem to encounter each other rather often in this newsgroup.
> Perhaps one day we'll meet in person. Where are you working these
> days? Still with Kenneth Rock?
>> I am also working with HeLa and would like to know its class I
> genotype. A former lab member told me that HeLa has been serotyped,
> by someone, on two separate occasions. As you might expect from
> serotyping, the results of the two tests did not quite agree. Test #1
> indicated the presence of the serotypes A3, A28, and B35. Test #2
> gave A68, B15, and B35.
>> I just searched GenBank using the key words "HeLa" and "HLA." I came
> up with the following IMAGE Consortium cDNA sequences from a
> Stratagene HeLa S3 cDNA library:
>> Accession #AA191477 and #AA191113: Both resemble Cw*1202. Clones may
> be full-length, but it does not appear that they were sequenced fully.
>> Accession #AA190717: Resembles Cw*1401. Again, it's short.
>> Accession #AA488534: Again incomplete, but resembles A*3402.
>> Accession #AA488401: Once more incomplete, but longer than the others.
> Best match is A*2601.
>> Some time next week, I will be visiting an Immunogenetics lab down at
> the Medical School here at Johns Hopkins. They have copies of the
> International Histocompatibility Workshop books. If someone has typed
> HeLa cells, maybe the information is there. Alternately -- is there
> someone reading this who has ready access to an IHW book? If you
> would share anything that the IHW might say about HeLa, that would be
> most helpful!
>> --
> John J. Ladasky Jr., Ph.D.
> Department of Biology
> Johns Hopkins University
> Baltimore MD 21218
> USA
> Earth
--
David R. Johnson, Ph.D.
Research Scientist, Department of Pathology
454 BCMM, 295 Congress Avenue
Yale University School of Medicine, New Haven, CT 06510 USA
Tel.: 203/737-2298, Fax: 203/737-2293
