While its possible the governments position on transmission of hepatitis C among boomers may have resulted in less testing, its critical today boomers forget any fears of stigma and get the easy blood test.
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Copyright © 1996
Blackwell Science
Detection of
hepatitis C virus (HCV)
proteins by
immunofluorescence and
HCV RNA genomic
sequences by
non-isotopic in situ
hybridization in bone
marrow and peripheral
blood mononuclear cells
of chronically
HCV-infected patients
D. SANSONNO, A. R. IACOBELLI, V. CORNACCHIULO, G. IODICE, and F. DAMMACCO
Department of Biomedical
Sciences and Human
Oncology, Section of
Internal Medicine and
Clinical Oncology,
University of Bari
Medical School, Bari,
Italy
Correspondence: Franco
Dammacco MD, Department
of Biomedical Sciences
and Human Oncology,
Section of Internal
Medicine and Clinical
Oncology, University of
Bari Medical School,
Policlinico, Piazza G.
Cesare, 11-70124 Bari,
Italy.
Accepted October 31,
1995.
Abstract
Immunofluorescence
(IF) to detect
HCV antigens and
non-isotopic
in situ
hybridization (NISH)
to detect HCV
RNA genome were
carried out on
bone marrow (BM)
and peripheral
blood (PB)
mononuclear
cells (MC) of 11
chronically
HCV-infected
patients. In
four patients
(36·4%) HCV
antigens were
detected in
monocytes/macrophages
as well as in B
lymphocytes in
both BMMC and
PBMC. Positive T
lymphocytes in
BMMC were found
in three of
them, but only
one patient
showed positive
T cells in PBMC.
NISH invariably
demonstrated
minus and plus
HCV RNA genomic
strands either
in monocytes/macrophages
or B and T
lymphocytes in
BMMC and PBMC in
the four HCV
antigen-positive
patients and in
two further
patients not
expressing viral
proteins in
blood MC. IF
signals appeared
diffusely
distributed
within the
cytoplasm, or as
brilliant
granules in
distinct
submembrane
areas or else in
cytoplasm
membrane. Nuclei
never stained.
Similarly, NISH
displayed HCV
RNA accumulation
restricted to MC
cytoplasm only,
nuclei being
persistently
negative. NISH,
however, was
unable to detect
cell membrane
signal.
Infection of
blood MC is a
common event in
naturally
acquired HCV
infection, since
none of these
patients was
conditioned by
immunomodulating
or
immunosuppressive
therapies. No
difference was
found in terms
of mean age,
length of
disease,
anti-HCV immune
response, type
and severity of
chronic liver
damage between
patients with
HCV-infected MC
and patients
without cell
infection. These
results
demonstrate that
HCV can infect
BMMC and PBMC
that represent
important
extrahepatic
sites of virus
replication, and
may help to
explain the
immunological
abnormalities
observed in
chronic HCV
carriers.
Full Text
The Full Text of
this article is
available as a
PDF (4.0M).
Selected
References
These references are
in PubMed. This may
not be the complete
list of references
from this article.
1.
Choo QL,
Weiner AJ,
Overby LR,
Kuo G,
Houghton M.
Hepatitis C
virus: the
major
causative
agent of
viral non-A,
non-B
hepatitis.
Br Med Bull.
1990;46:42341.
[PubMed]
2.
Colombo M,
Choo QL, Del
Ninno E, et
al.
Prevalence
of
antibodies
to hepatitis
C virus in
Italian
patients
with
hepatocellular
carcinoma.
Lancet.
1989;2:10068.
[PubMed]
3.
Gerber MA,
Shieh YSC,
Shim K-S, et
al.
Detection of
replicative
hepatitis C
virus
sequences in
hepatocellular
carcinoma.
Am J Pathol.
1992;141:12717.
[PMC
free article]
[PubMed]
4.
Paterlini P,
Driss F,
Nalpas B,
Pisi E,
Franco D,
Berthelot P,
Brechot C.
Persistence
of hepatitis
B and
hepatitis C
viral
genomes in
primary
liver
cancers from
HBsAg-negative
patients: a
study of
low-endemic
area.
Hepatology.
1993;17:2029.
[PubMed]
5.
Houghton M,
Weiner A,
Han J, Kuo
G, Choo QL.
Molecular
biology of
the
hepatitis C
viruses:
implications
for
diagnosis,
development
and control
of viral
disease.
Hepatology.
1991;14:3818.
[PubMed]
6.
Choo QL,
Richman KH,
Han JH, et
al. Genetic
organization
and
diversity of
the
hepatitis C
virus.
Proc Natl
Acad Sci
USA.
1991;88:24515.
[PMC
free article]
[PubMed]
7.
Dienstag JL.
Hepatitis
non-A,
non-B: C at
last.
Gastroenterology.
1990;99:117780.
[PubMed]
8.
Erickson AL,
Houghton M,
Choo QL,
Weiner AJ,
Ralston R,
Muchmore E,
Walker CM.
Hepatitis C
virus-specific
CTL
responses in
the liver of
chimpanzees
with acute
and chronic
hepatitis C.
J Immunol.
1993;151:418999.
[PubMed]
9.
Minutello
AM, Pileri
P, Unutmatz
D, et al.
Compartmentalization
of T
lymphocytes
to the site
of disease:
intrahepatic
CD4+
T cells,
specific for
the protein
NS4 of
hepatitis C
virus in
patients
with chronic
hepatitis C.
J Exp Med.
1993;178:1725.
[PMC
free article]
[PubMed]
10.
Ferrari C,
Valli A,
Galati L, et
al. T-cell
response to
structural
and
nonstructural
hepatitis C
virus
infections.
Hepatology.
1994;19:28695.
[PubMed]
11.
Alter HJ. To
C or not to
C: these are
the
questions.
Blood.
1995;85:168195.
[PubMed]
12.
Krawczynski
K, Beach MJ,
Bradley DW,
et al.
Hepatitis C
virus
antigen in
hepatocytes:
immunomorphologic
detection
and
identification.
Gastroenterology.
1992;103:6229.
[PubMed]
13.
Hiramatsu N,
Hayashi N,
Haruna Y, et
al.
Immunohistochemical
detection of
hepatitis C
virus-infected
hepatocytes
in chronic
liver
disease with
monoclonal
antibodies
to core,
envelope and
NS3 regions
of the
hepatitis C
virus
genome.
Hepatology.
1992;16:30611.
[PubMed]
14.
Sansonno D,
Dammacco F.
Hepatitis C
virus c100
antigen in
liver tissue
from
patients
with acute
and chronic
infection.
Hepatology.
1993;18:2405.
[PubMed]
15.
Blight K,
Lesniewski
RR, Labrooy
JT, Gowans
EJ.
Detection
and
distribution
of hepatitis
C-specific
antigens in
naturally
infected
liver.
Hepatology.
1994;20:5537.
[PubMed]
16.
Negro F,
Pacchioni D,
Shimizu Y,
Miller RH,
Bussolati G,
Purcell RH,
Bonino F.
Detection of
intrahepatic
replication
of hepatitis
C virus RNA
by in
situ
hybridization
and
comparison
with
histopathology.
Proc Natl
Acad Sci
USA.
1992;89:224751.
[PMC
free article]
[PubMed]
17.
Yamada S,
Kojl T,
Nozawa M,
Kiyosama K,
Nakane PK.
Detection of
hepatitis C
virus (HCV)
RNA in
paraffin
embedded
tissue
sections of
human liver
of non-A,
non-B
hepatitis
patients by
in situ
hybridization.
J Clin Lab
Anal.
1992;6:4046.
[PubMed]
18.
Tigges MA,
Koelle D,
Hartog K,
Sekulovich
RE, Corey L,
Burke RL.
Human CD8+
herpes
simplex
virus-specific
cytotoxic
T-lymphocyte
clones
recognize
diverse
virion
protein
antigens.
J Virol.
1992;66:162234.
[PMC
free article]
[PubMed]
19.
Oldstone
MBA. Viral
persistence.
Cell.
1989;56:51720.
[PubMed]
20.
Lamelin JP,
Zoulim F,
Trepo C.
Lymphotropism
of hepatitis
B and C
virus: an
update and a
newcomer.
Int J Clin
Lab Res.
1995;25:16.
[PubMed]
21.
Bouffard P,
Hayashi PH,
Acevedo R,
Levy N,
Zeldis JB.
Hepatitis C
virus is
detected in
a
monocyte/macrophage
subpopulation
of
peripheral
blood
mononuclear
cells of
infected
patients.
J Infect
Dis.
1992;166:127680.
[PubMed]
22.
Zignego AL,
Macchia D,
Monti M, et
al.
Infection of
peripheral
mononuclear
blood cells
by hepatitis
C virus.
J Hepatol.
1992;15:3826.
[PubMed]
23.
Bartolomθ J,
Castillo I,
Quiroga JA,
Navas S,
Carreno V.
Detection of
hepatitis C
virus RNA in
serum and
peripheral
blood
mononuclear
cells.
J Hepatol.
1993;17:s90s93.
[PubMed]
24.
Ferri C,
Monti M, La
Civita L, et
al.
Infection of
peripheral
blood-mononuclear
cells by
hepatitis C
virus in
mixed
cryoglobulinemia.
Blood.
1993;82:37014.
[PubMed]
25.
Gabrielli A,
Manzin A,
Candela M,
Caniglia ML,
Paolucci S,
Danieli MG,
Clementi M.
Active
hepatitis C
virus
infection in
bone marrow
and
peripheral
blood
mononuclear
cells from
patients
with mixed
cryoglobulinemia.
Clin Exp
Immunol.
1994;97:8793.
[PMC
free article]
[PubMed]
26.
Moldvay J,
Deny P, Pol
S, Brechot
C, Lamas E.
Detection of
hepatitis C
virus RNA in
peripheral
blood
mononuclear
cells of
infected
patients by
in situ
hybridization.
Blood.
1994;83:26973.
[PubMed]
27.
Gutierrez C,
Bernabe RR,
Vega J,
Kreisler M.
Purification
of human T
and B cells
by a
discontinuous
density
gradient of
Percoll.
J Immunol
Methods.
1979;29:5763.
[PubMed]
28.
Houghton M,
Choo QL, Kuo
G. European
Patent
Application.
Publication
no. 0318216
and no.
0388232.
29.
Yamada G,
Nishimoto H,
Endou H, et
al.
Localization
of hepatitis
C viral RNA
and capsid
protein in
human liver.
Dig Dis Sci.
1993;38:8827.
[PubMed]
30.
Schmitz GG,
Walter T,
Seibl R,
Kessler C.
Nonradioactive
labeling of
oligonucleotides
in vitro
with the
hapten
digoxigenin
by tailing
with
terminal
transferase.
Anal
Biochem.
1991;192:22231.
[PubMed]
31.
Eggerding
FA, Peters
J, Lee RK,
Inderlied
CB.
Detection of
rubella
virus gene
sequences by
enzymatic
amplification
and direct
sequencing
of amplified
DNA.
J Clin
Microbiol.
1991;29:94552.
[PMC
free article]
[PubMed]
32.
Dammacco F,
Sansonno D,
Beardsley A,
Gowans EJ.
Failure to
detect
hepatitis C
virus (HCV)
genome by
polymerase
chain
reaction in
human
anti-HCV-positive
intravenous
immunoglobulins.
Clin Exp
Immunol.
1993;92:20510.
[PMC
free article]
[PubMed]
33.
Gudat F,
Bianchi L.
Evidence for
phasic
sequences in
nuclear
HBcAg
formation
and cell
membrane-directed
flow of core
particles in
chronic
hepatitis B.
Gastroenterology.
1977;73:11947.
[PubMed]
34.
Cleaves GR,
Ryan TE,
Schlesinger
RW.
Identification
and
characterization
of type 2
Dengue virus
replicative
form RNAs.
Virology.
1981;111:7383.
[PubMed]
35.
Halstead SB,
O'Rourke EJ.
Antibody-enhanced
Dengue virus
infection in
primate
leukocytes.
Nature.
1977;265:73974.
[PubMed]
36.
Peiris JSM,
Porterfield
JS.
Antibody-mediated
enhancement
of
Flavivirus
replication
in
macrophage-like
cell lines.
Nature.
1979;282:50911.
[PubMed]
37.
Porterfield
JS.
Antibody-dependent
enhancement
of viral
infectivity.
In:
Maramorosch
K, Murphy
FA, Shatkin
AJ, editors.
Advances in
virus
research.
Vol. 31.
London:
Academic
Press Inc;
1986. pp.
33555.
38.
Kuo G, Choo
QL, Alter
HJ, et al.
An assay for
circulating
antibodies
to a major
etiologic
virus of
human non-A,
non-B
hepatitis.
Science.
1989;244:3624.
[PubMed]
39.
Sansonno D,
Dammacco F.
Antibodies
to hepatitis
C virus in
non-A, non-B
post
transfusion
and
cryptogenetic
chronic
liver
disease.
Lancet.
1989;ii:7989.
[PubMed]
40.
Agnello V,
Chung RT,
Kaplan LM. A
role for
hepatitis C
virus
infection in
type II
cryoglobulinemia.
N Engl J
Med.
1992;327:14905.
[PubMed]
41.
Bukh J,
Purcell RH,
Miller RH.
At least 12
genotypes of
hepatitis C
virus
predicted by
sequence
analysis of
the putative
E1 gene of
isolates
collected
worldwide.
Proc Natl
Acad Sci
USA.
1993;90:82348.
[PMC
free article]
[PubMed]
42.
Sansonno D,
Procaccio P,
Dammacco F.
Kinetics of
anti-hepatitis
C virus
response
during acute
hepatitis in
an
immunosuppressed
HCV-chronically
infected
patient.
Digestion.
1991;50:5260.
[PubMed]
43.
Fan FS,
Tzeng CH,
Hsiao KI, Hu
ST, Liu WT,
Chen PM.
Withdrawal
of
immunosuppressive
therapy in
allogenic
bone marrow
transplantation
reactivates
chronic
viral
hepatitis C.
Bone Marrow
Transplant.
1991;8:41720.
[PubMed]
44.
Dammacco F,
Sansonno D.
Antibodies
to hepatitis
C virus in
essential
mixed
cryoglobulinemia.
Clin Exp
Immunol.
1992;87:3526.
[PMC
free article]
[PubMed]
45.
Lunel F,
Musset L,
Franjeul L,
et al.
Cryoglobulinemia
in chronic
liver
diseases:
role of
hepatitis C
virus and
liver
damage.
Gastroenterology.
1994;106:1291300.
[PubMed]
46.
Lunel F.
Hepatitis C
virus and
autoimmunity:
fortuitous
association
or reality?
Gastroenterology.
1994;107:15505.
[PubMed]
47.
Sansonno D,
Cornacchiulo
V, Iacobelli
AR, Di
Stefano R,
Lospalluti
M, Dammacco
F.
Localization
of hepatitis
C virus
(HCV)
antigens in
liver and
skin tissues
of
chronically
HCV-infected
patients
with mixed
cryoglobulinemia.
Hepatology.
1995;21:30512.
[PubMed]
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