Discrimination between antibodies against HTLV-I, HTLV-II or...

Details Discrimination between antibodies against HTLV-I, HTLV-II or... Discrimination between antibodies against HTLV-I, HTLV-II or...

1991-10-07

2001-06-05

Stucker, Jeffery (Department: 1648)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving virus or bacteriophage

C435S007100, C435S974000, C435S975000, C530S324000, C530S325000, C530S326000, C530S328000, C530S826000, C530S327000

Reexamination Certificate

active

06242174

ABSTRACT:

The present invention relates to a method of discriminating between specific antibodies in samples of sera or other body fluids from humans or other primates containing antibodies arising from infection with HTLV-I, HTLV-II or related retroviruses. Additionally, it relates to an immunoassay kit adapted for said method of discrimination, and new peptides and a method of detecting antibodies with said peptides.
BACKGROUND
Up to now the following techniques for differentiating infection with the two viruses have been used: Virus isolation with typing, serological techniques (based on antibody competition or neutralization), or nucleic acid techniques (nucleic acid amplification or hybridization). Most of these techniques are laborious and require special competence.
Human T-lymphotropic virus type I (HTLV-I) and type II (HTLV-II) are widespread human retroviruses (a short review is given in ref. 6) (1, 2, 3, 20). HTLV-II has for several years been considered to be rare, but has recently proved to be a rather common infection among intravenous drug abusers primarily in the United States of America. The viruses cross-react serologically. It is therefore impossible to discriminate between an infection with one virus from an infection with the other with current antibody tests. It may prove clinically important to differentiate between infections with the two viruses. HTLV-I is associated with a type of leukemia (Adult T cell Leukemia; ATL) while HTLV-II has been observed in a few cases of hairy cell leukemia. There is a need for simple tests to differentiate between the two infections.
Even if the amino acid sequences of HTLV-I and HTLV-II proteins are similar there are several regions where they are markedly different. Our idea is to use synthetic peptides from such regions as antigens in antibody tests. We have found peptides with sequences which e.g. are suitable for solid phase immunoassays and which give a type-specific antibody reactivity. We have found techniques where we use them to discern infection with HTLV-I from infection with HTLV-II.
DESCRIPTION OF THE INVENTION
One aspect of the invention is directed to a method of discriminating between specific antibodies in samples of sera or other body fluids from humans or other primates containing antibodies arising from infection with HTLV-I, containing antibodies arising from infection with HTLV-II or containing antibodies arising from infection with related retroviruses, whereby the sample to be analyzed is subjected to at least four immunoassays, each using a different diagnostic antigen selected from the following groups a) to d):
a) peptides comprising a sequence of at least 17 amino acid residues which corresponds to a sequence of HTLV-I gag comprising antigenic structures;
b) peptides comprising a sequence of at least 17 amino acid residues which corresponds to a sequence of HTLV-II gag comprising antigenic structures;
c) peptides comprising a sequence of at least 17 amino acid residues which corresponds to a sequence of HTLV-I env comprising antigenic structures;
d) peptides comprising a sequence of at least 17 amino acid residues which corresponds to a sequence of HTLV-II env comprising antigenic structures;
with the proviso that at least one peptide from each of the groups a) to d) is selected and, further, that at least one pair of peptides corresponding to at least partially overlapping sequences of HTLV-I and HTLV-II is selected from each of the groupages a) plus b), and c) plus d),
and that the analyzed, different binding strengths of the antibodies of the sample in said at least four immunoassays are used to discriminate between antibodies arising from infection with one specific retrovirus and antibodies arising from infection with other specific retroviruses.
In an embodiment of this aspect of the invention the diagnostic antigens are selected in the above manner from the peptides:
a) HTLV-I
gag 130-197
PVMHPHGAPPNHRPWQMKDLQAIKQE-
VSQAAPGSPQFMQTIRLAVQQFDPTA-
KDLQDLLQYLCSSLVA
b) HTLV-I
gag 137-214
PILHPPGAPSAHRPWQMKDLQAIKQEV-
SSSALGSPQFMQTLRLAVQQFDPTAKD-
LQDLLQYLCSSLVV
a) HTLV-I
gag 298-349
LRSLAYSNANKECQKLLQARGHTNSPL-
GDMLRACQTWTPKDKTKVLVVQPKK
b) HTLV-II
gag 305-356
LRSLAYSNANKECQKILQARGHTNSPL-
GEMLRTCQAWTPKDKTKVLVVQPRR
a) HTLV-I
gag 4-20
IFSRSASPIPRPPRGLA
b) HTLV-II
gag 4-20
IHGLSPTPIPKAPRGLS
a) HTLV-I
gag 111-130
PDSDPQIPPPYVEPTAPQVL
b) HTLV-II
gag 117-136
PSPEAHVPPPYVEPTTTQCP
a) HTLV-I
gag 265-285
SILQGLEEPYHAFVERLNIAL
a) HTLV-I
gag 302-320
LAYSNANKECQKLLQARGH
a) HTLV-I
gag 323-341
SPLGDMLRACQTWTPKDKT
a) HTLV-I
gag 337-355
PKDKTKVLVVQPKKPPPNQ
b) HTLV-II
gag 343-361
PKDKTKVLVVQPRRPPPTQ
a) HTLV-I
gag 378-399
PCPLCQDPTHWKRDCPRLKPT
a) HTLV-I
gag 392-411
DCPRLKPTIPEPEPEEDALL
b) HTLV-II
gag 398-416
DCPQLKPPQEEGEPLLLDL
c) HTLV-I
env 190-213
LLPHSNLDHILEPSIPWKSKLLTL
d) HTLV-II
env 186-209
LVHDSDLEHVLTPSTSWTTKILKF
c) HTLV-I
env 290-312
HNSLILPPFSLSPVPTLGSRSRR
c) HTLV-I
env 360-378
AIVKNHKNLLKIAQYAAQN
c) HTLV-I
env 376-392
AQNRRGLDLLFWEQGGL
c) HTLV-I
env 380-398
RGLDLLFWEQGGLCKALQE
c) HTLV-I
env 465-488
RQLRHLPSRVRYPHYSLILPESSL
d) HTLV-II
env 463-486
IQALPQRLQNRHNQYSLINPETML
In a preferred embodiment at least the following peptides are selected:
a) HTLV-I
gag 111-130
PDSDPQIPPPYVEPTAPQVL
b) HTLV-II
gag 117-136
PSPEAHVPPPYVEPTTTQCP
c) HTLV-I
env 190-213
LLPHSNLDHILEPSIPWKSKLLTL
d) HTLV-II
env 185-209
LVHDSDLEHVLTPSTSWTTKILKF
In a further preferred embodiment the sample to be analyzed is subjected to at least eight immunoassays and the analyzed pattern of binding strengths is processed with a computer program.
Optionally, at least one of the selected peptides is attached to an inert soluble or insoluble carrier.
Another aspect of the invention is directed to a peptide, which corresponds to a sequence of HTLV-I, HTLV-II or a related retrovirus each comprising antigenic structures and which comprises a sequence of at least 17 amino acid residues selected from the following sequences:
HTLV-I
gag 130-197
PVMHPHGAPPNHRPWQMKDLQAIKQE-
VSQAAPGSPQFMQTIRLAVQQFDPTA-
KDLQDLLQYLCSSLVA
HTLV-I
gag 137-214
PILHPPGAPSAHRPWQMKDLQAIKQEV-
SSSALGSPQFMQTLRLAVQQFDPTAKD-
LQDLLQYLCSSLVV
HTLV-I
gag 298-349
LRSLAYSNANKECQKLLQARGHTNSPL-
GDMLRACQTWTPKDKTKVLVVQPKK
HTLV-II
gag 305-356
LRSLAYSNANKECQKILQARGHTNSPL-
GEMLRTCQTWAPKDKTKVLVVQPRR
HTLV-I
gag 4-20
IFSRSASPIPRPPRGLA
HTLV-II
gag 4-20
IHGLSPTPIPKAPRGLS
HTLV-I
gag 111-130
PDSDPQIPPPYVEPTAPQVL
HTLV-II
gag 117-136
PSPEAHVPPPYVEPTTTQCP
HTLV-I
gag 265-285
SILQGLEEPYHAFVERLNIAL
HTLV-I
gag 302-320
LAYSNANKECQKLLQARGH
HTLV-I
gag 323-341
SPLGDMLRACQTWTPKDKT
HTLV-I
gag 337-355
PKDKTKVLVVQPKKPPPNQ
HTLV-II
gag 343-361
PKDKTKVLVVQPRRPPPTQ
HTLV-I
gag 378-399
PCPLCQDPTHWKRDCPRLKPT
HTLV-I
gag 392-411
DCPRLKPTIPEPEPEEDALL
HTLV-II
gag 398-416
DCPQLKPPQEEGEPLLLDL
HTLV-I
env 190-213
LLPHSNLDHILEPSIPWKSKLLTL
HTLV-II
env 186-209
LVHDSDLEHVLTPSTSWTTKILKF
HTLV-I
env 290-312
HNSLILPPFSLSPVPTLGSRSRR
HTLV-I
env 360-378
AIVKNHKNLLKIAQYAAQN
HTLV-I
env 376-392
AQNRRGLDLLFWEQGGL
HTLV-I
env 380-398
RGLDLLFWEQGGLCKALQE
HTLV-I
env 465-488
RQLRHLPSRVRYPHYSLILPESSL
HTLV-II
env 463-486
IQALPQRLQNRHNQYSLINPETML
Yet another aspect of the invention is directed to a method of detecting antibodies arising from infection with HTLV-I, HTLV-II or a related retrovirus in a sample of serum or other body fluid from a human or an other primate, whereby said sample is subjected to an immunoassay using as a diagnostic antigen at least one peptide of the invention.
Still another aspect of the invention is directed to an immunoassay kit for the discrimination between samples of sera or other body fluids from humans or other primates containing antibodies arising from infection with HTLV-I, containing antibodies arising from infection with HTLV-II or containing antibodies arising from infection with related retroviruses, which kit comprises at least four peptides selected from the following groups a) to d):
a) peptides comprising a sequence of at least 17 amino acid residues which corresponds to a sequence of HTLV-I gag comprising antigenic structures;
b) peptides comprising a sequence of at least 17 amino acid residues which corresponds to a sequ

Affiliated with

Also associated with

Rating

Discrimination between antibodies against HTLV-I, HTLV-II or... does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Discrimination between antibodies against HTLV-I, HTLV-II or..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Discrimination between antibodies against HTLV-I, HTLV-II or... will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2533624