WU Polyomavirus in Patients Infected with HIV or Hepatitis C Virus, Connecticut, USA, 2007

[Guest guest]

http://www.cdc.gov/eid/content/15/7/1095.htm

EID Journal Home> Volume 15, Number 7–July 2009

Volume 15, Number 7–July 2009

Dispatch

WU Polyomavirus in Patients Infected with HIV or Hepatitis C Virus, Connecticut,

USA, 2007

A. , Carla Weibel, Ferguson, Marie L. Landry, and S.

Kahn

Author affiliation: Yale University School of Medicine, New Haven, Connecticut,

USA

Abstract

WU polyomavirus (WUPyV) was detected in 10 (8.3%) of 121 HIV-positive plasma

specimens, 0 (0%) of 120 HIV-negative serum specimens, and 2 (2.5%) of 79

hepatitis C virus (HCV)–positive serum specimens. KI polyomavirus was not

detected in HIV-positive plasma or HCV-positive serum specimens. HIV-infected

persons may be susceptible to systemic WUPyV infection.

In 2007, 2 new human polyomaviruses, KI polyomavirus (KIPyV) and WU polyomavirus

(WUPyV), were identified. KIPyV was initially detected in an extract obtained

from 20 pooled randomly selected nasopharyngeal aspirates, and WUPyV was

detected in a nasopharyngeal aspirate from a 3-year-old child from Australia who

had a diagnosis of pneumonia (1,2). These viruses have since been detected in

respiratory tract specimens from symptomatic and asymptomatic children, although

no clear association with respiratory disease has been demonstrated (3–6).

Previously identified human polyomaviruses (BK virus [bKV] and JC virus [JCV])

cause clinical disease in immunocompromised persons (7,8). Although viremia may

be associated with immunosuppression, correlation of JCV DNA in peripheral blood

with development of progressive multifocal encephalopathy in AIDS patients

remains controversial (9). BKV DNA has been detected in blood of renal

transplant patients, and BKV load may be predictive of polyomavirus-associated

nephropathy (10).

The Study

The pathogenesis and clinical spectra of WUPyV and KIPyV, particularly in

immunocompromised persons, have not been defined. To investigate whether WUPyV

or KIPyV is present in persons with chronic viral infection and perhaps

compromised immunity, we conducted a cross-sectional study in which we screened

the following for WUPyV and KIPYV DNA: plasma samples from HIV-infected persons,

serum samples from hepatitis C virus (HCV)–infected persons, and a control group

of HIV-negative persons.

Three groups of samples submitted to the Clinical Virology Laboratory at

Yale–New Haven Hospital in 2007 were screened: HIV PCR-positive plasma, HCV

PCR-positive serum, and HIV antibody-negative serum. Patient identifiers were

removed and these specimens were tested as part of our ongoing investigation for

newly identified viruses. Collection of specimens and clinical data was approved

by the Yale University Human Investigation Committee and was compliant with

Health Insurance Portability and Accountability Act regulations.

Nucleic acids were extracted from each specimen by using QIAamp nucleic acid

purification kits (QIAGEN, Valencia, CA, USA). Screening for WUPyV DNA has been

described (6). Briefly, we performed an initial PCR screening specific for the

virus capsid protein 2 (VP2) gene by using primers described by Gaynor et al.

(2) and a nested PCR (Table 1, primers 1 and 2). To confirm results, DNA from

all PCR-positive samples was reextracted and screened with primers specific for

the region of the genome containing the noncoding control region (NCCR), which

includes the virus origin of replication (genome coordinates 5213 to nt 36 of

the circular viral genome). This screening included an initial PCR (Table 1,

primers 3 and 4) and a nested PCR (Table 1, primers 5 and 6). The nested PCR

generated a 328-bp amplicon. Screening for KIPyV DNA by PCR included a nested

PCR specific for the VP1 gene according to the protocol described by Allander et

al. (1). Positive and negative controls were included in each set of PCRs.

All PCR products were sequenced by using 377 DNA automated sequencers (Applied

Biosystems, City, CA, USA) at the W.M. Keck Biotechnology Resource

Laboratory at Yale University School of Medicine. For WUPyV, phylogenetic

analysis was performed on a 194-bp fragment within the amplified region of NCCR

(nt 5197 to nt 159 of the circular viral genome) by using Lasergene MegAlign

software (DNASTAR Inc., Madison, WI, USA) (ClustalW alignment method). The only

clinical data available for these deidentified serum specimens were HIV/HCV

status and virus loads. HIV and HCV virus loads were determined in the Clinical

Virology Laboratory by quantitative reverse transcription–PCR using commercially

available diagnostic tests. The Fisher exact test was used to determine whether

the difference in the percentage of WUPyV-positive specimens in HIV-positive and

HIV-negative patients was statistically significant.

Ten (8.3%) of 121 HIV-positive specimens and 0 (0%) of 120 HIV-negative samples

were positive for WUPyV (p85% of the persons screened will have detectable

antibodies to WUPyV (11). Whether antibody status for WUPyV correlates with

viremia is unknown. Viremia may represent primary infection or reactivation of

latent infection.

Viremia has been described for JCV and BKV. JCV DNA in serum/plasma may

correlate with the degree of immunosuppression. However, blood from viremic

persons infected with JCV has a low positive predictive value for development of

progressive multifocal encephalopathy in AIDS patients (9). A recent study

suggested that screening for BKV replication is useful in identifying patients

at risk for BKV-associated nephropathy, which may enable early interventions

such as renal biopsy and reduction of immunosuppression (12). However, because

of the lack of clinical data available for WUPyV-positive persons in our study,

it was not possible to make any clinical correlations.

The absence of KIPyV in HIV-positive or HCV-positive peripheral blood specimens

suggests that host susceptibility for KIPyV may differ from that of WUPyV, as

for JCV and BKV. However, this hypothesis was not supported by a recent study

that reported KIPyV and WUPyV in autopsy lymphoid tissues of AIDS patients (13).

Whether WUPyV or KIPyV cause disease in HIV-positive persons or other

populations remains to be determined.

Our data demonstrate that WUPyV was detected in peripheral blood of HIV- and

HCV-infected persons. However, the scope of this study was limited because

clinical data were not available for study participants. Whether WUPyV or KIPyV

have oncogenicity or other pathogenicity in immunocompromised hosts remain to be

determined. The role of polyomaviruses in human cancers has been extensively

investigated but conclusive evidence is lacking (14). The genome of Merkel cell

polyomavirus, a new polyomavirus, was found to be integrated within the cellular

genome of Merkel cell carcinoma tissue samples, which suggests a role for this

virus in a specific tumor (15). Therefore, studies to assess the oncogenic

potential of WUPyV and KIPyV are also needed.

Acknowledgments

We thank for continued support, intellectual and scientific input,

and exchange and critical review of the data; and the staff of the Clinical

Virology Laboratory, Yale–New Haven Hospital, for assistance in this study.

This study was supported by National Institutes of Health grant T32 A107210-20

and a MedImmune pediatric fellowship grant.

Dr is a fellow in Pediatric Infectious Diseases at Yale University School

of Medicine. His research interests are the biology, epidemiology, and oncogenic

potential of human polyomaviruses.

References

Allander T, sson K, Gupta S, Bjerkner A, Gordana B, Perrson MA, et al.

Identification of a third human polyomavirus. J Virol. 2007;81:4130–6. PubMed

DOI

Gaynor AM, Nissen MD, Whiley DM, MacKay IM, Lambert SB, Wu G, et al.

Identification of a novel polyomavirus from patients with acute respiratory

tract infections. PLoS Pathog. 2007;3:e64. PubMed DOI

Norja P, Ubillos I, Templeton K, Simmonds P. No evidence for an association

between infections with WU and KI polyomaviruses and respiratory disease. J Clin

Virol. 2007;40:307–11. PubMed DOI

Le BM, Demertzis LM, Wu G, Tibbets RJ, Buller R, Arens MQ, et al. Clinical and

epidemiologic characterization of WU polyomavirus infection, St. Louis,

Missouri. Emerg Infect Dis. 2007;13:1936–8.

Bialasiewicz S, Whiley DM, Lambert SB, K, Bletchly C, Wang D, et al. A

newly reported human polyomavirus, KI virus, is present in the respiratory tract

of Australian children. J Clin Virol. 2007;40:15–8. PubMed DOI

Wattier RL, Vazquez MV, Weibel C, Shapiro ED, Ferguson D, Landry ML, et al. Role

of human polyomaviruses in respiratory tract disease in young children. Emerg

Infect Dis. 2008;14:1766–8. PubMed DOI

Khalili K, Gordon J, White MK. The polyomavirus, JCV and its involvement in

human disease. Adv Exp Med Biol. 2006;577:274–87. PubMed DOI

Hirsch HH. Polyomavirus BK nephropathy: a (re-)emerging complication in renal

transplantation. Am J Transplant. 2002;2:25–30. PubMed DOI

Andreoletti L, Lescieux A, Lambert B, Si-Mohamed A, Matta M, Wattre P, et al.

Semiquantitative detection of JCV-DNA in peripheral blood leukocytes from

HIV-1-infected patients with or without progressive multifocal

leukoencephalopathy. J Med Virol. 2002;66:1–7. PubMed DOI

Hymes LC, Warshaw BL. Polyomavirus (BK) in pediatric renal transplants:

evaluation of viremic patients with and without BK associated nephritis. Pediatr

Transplant. 2006;10:920–2. PubMed DOI

Stolt A, Sasnauskas K, Koskela P, Lehtinen M, Dillner J. Seroepidemiology of the

human polyomaviruses. J Gen Virol. 2003;84:1499–504. PubMed DOI

Costa C, Bergallo M, Astegiano S, Terlizzi ME, Sidoti F, Segoloni GP, et al.

Monitoring of BK virus replication in the first year following renal

transplantation. Nephrol Dial Transplant. 2008;23:3333–6. PubMed DOI

Sharp CP, Norja P, J, Bell JE, Simmonds P. Reactivation and mutation of

newly discovered WU, KI, and Merkel cell carcinoma polyomaviruses in

immunosuppressed individuals. J Infect Dis. 2009;199:398–404. PubMed DOI

zur Hausen H. Novel human polyomaviruses–re-emergence of a well known virus

family as possible human carcinogens. Int J Cancer. 2008;123:247–50. PubMed DOI

Feng H, Shuda M, Chang Y, PS. Clonal integration of a polyomavirus in

human Merkel cell carcinoma. Science. 2008;319:1096–100. PubMed DOI

Suggested Citation for this Article

MA, Weibel C, Ferguson D, Landry ML, Kahn JS. WU polyomavirus in patients

infected with HIV or hepatitis C virus, Connecticut, USA, 2007. Emerg Infect Dis

[serial on the Internet]. 2009 Jul [date cited]. Available from

http://www.cdc.gov/EID/content/15/7/1095.htm

DOI: 10.3201/eid1507.090150

S. Kahn, Department of Pediatrics, Division of Infectious Diseases, Yale

University School of Medicine, PO Box 208064, New Haven, CT 06520-8064, USA;

email: jeffrey.kahn@...

[Guest guest]

http://www.cdc.gov/eid/content/15/7/1095.htm

EID Journal Home> Volume 15, Number 7–July 2009

Volume 15, Number 7–July 2009

Dispatch

WU Polyomavirus in Patients Infected with HIV or Hepatitis C Virus, Connecticut,

USA, 2007

A. , Carla Weibel, Ferguson, Marie L. Landry, and S.

Kahn

Author affiliation: Yale University School of Medicine, New Haven, Connecticut,

USA

Abstract

WU polyomavirus (WUPyV) was detected in 10 (8.3%) of 121 HIV-positive plasma

specimens, 0 (0%) of 120 HIV-negative serum specimens, and 2 (2.5%) of 79

hepatitis C virus (HCV)–positive serum specimens. KI polyomavirus was not

detected in HIV-positive plasma or HCV-positive serum specimens. HIV-infected

persons may be susceptible to systemic WUPyV infection.

In 2007, 2 new human polyomaviruses, KI polyomavirus (KIPyV) and WU polyomavirus

(WUPyV), were identified. KIPyV was initially detected in an extract obtained

from 20 pooled randomly selected nasopharyngeal aspirates, and WUPyV was

detected in a nasopharyngeal aspirate from a 3-year-old child from Australia who

had a diagnosis of pneumonia (1,2). These viruses have since been detected in

respiratory tract specimens from symptomatic and asymptomatic children, although

no clear association with respiratory disease has been demonstrated (3–6).

Previously identified human polyomaviruses (BK virus [bKV] and JC virus [JCV])

cause clinical disease in immunocompromised persons (7,8). Although viremia may

be associated with immunosuppression, correlation of JCV DNA in peripheral blood

with development of progressive multifocal encephalopathy in AIDS patients

remains controversial (9). BKV DNA has been detected in blood of renal

transplant patients, and BKV load may be predictive of polyomavirus-associated

nephropathy (10).

The Study

The pathogenesis and clinical spectra of WUPyV and KIPyV, particularly in

immunocompromised persons, have not been defined. To investigate whether WUPyV

or KIPyV is present in persons with chronic viral infection and perhaps

compromised immunity, we conducted a cross-sectional study in which we screened

the following for WUPyV and KIPYV DNA: plasma samples from HIV-infected persons,

serum samples from hepatitis C virus (HCV)–infected persons, and a control group

of HIV-negative persons.

Three groups of samples submitted to the Clinical Virology Laboratory at

Yale–New Haven Hospital in 2007 were screened: HIV PCR-positive plasma, HCV

PCR-positive serum, and HIV antibody-negative serum. Patient identifiers were

removed and these specimens were tested as part of our ongoing investigation for

newly identified viruses. Collection of specimens and clinical data was approved

by the Yale University Human Investigation Committee and was compliant with

Health Insurance Portability and Accountability Act regulations.

Nucleic acids were extracted from each specimen by using QIAamp nucleic acid

purification kits (QIAGEN, Valencia, CA, USA). Screening for WUPyV DNA has been

described (6). Briefly, we performed an initial PCR screening specific for the

virus capsid protein 2 (VP2) gene by using primers described by Gaynor et al.

(2) and a nested PCR (Table 1, primers 1 and 2). To confirm results, DNA from

all PCR-positive samples was reextracted and screened with primers specific for

the region of the genome containing the noncoding control region (NCCR), which

includes the virus origin of replication (genome coordinates 5213 to nt 36 of

the circular viral genome). This screening included an initial PCR (Table 1,

primers 3 and 4) and a nested PCR (Table 1, primers 5 and 6). The nested PCR

generated a 328-bp amplicon. Screening for KIPyV DNA by PCR included a nested

PCR specific for the VP1 gene according to the protocol described by Allander et

al. (1). Positive and negative controls were included in each set of PCRs.

All PCR products were sequenced by using 377 DNA automated sequencers (Applied

Biosystems, City, CA, USA) at the W.M. Keck Biotechnology Resource

Laboratory at Yale University School of Medicine. For WUPyV, phylogenetic

analysis was performed on a 194-bp fragment within the amplified region of NCCR

(nt 5197 to nt 159 of the circular viral genome) by using Lasergene MegAlign

software (DNASTAR Inc., Madison, WI, USA) (ClustalW alignment method). The only

clinical data available for these deidentified serum specimens were HIV/HCV

status and virus loads. HIV and HCV virus loads were determined in the Clinical

Virology Laboratory by quantitative reverse transcription–PCR using commercially

available diagnostic tests. The Fisher exact test was used to determine whether

the difference in the percentage of WUPyV-positive specimens in HIV-positive and

HIV-negative patients was statistically significant.

Ten (8.3%) of 121 HIV-positive specimens and 0 (0%) of 120 HIV-negative samples

were positive for WUPyV (p85% of the persons screened will have detectable

antibodies to WUPyV (11). Whether antibody status for WUPyV correlates with

viremia is unknown. Viremia may represent primary infection or reactivation of

latent infection.

Viremia has been described for JCV and BKV. JCV DNA in serum/plasma may

correlate with the degree of immunosuppression. However, blood from viremic

persons infected with JCV has a low positive predictive value for development of

progressive multifocal encephalopathy in AIDS patients (9). A recent study

suggested that screening for BKV replication is useful in identifying patients

at risk for BKV-associated nephropathy, which may enable early interventions

such as renal biopsy and reduction of immunosuppression (12). However, because

of the lack of clinical data available for WUPyV-positive persons in our study,

it was not possible to make any clinical correlations.

The absence of KIPyV in HIV-positive or HCV-positive peripheral blood specimens

suggests that host susceptibility for KIPyV may differ from that of WUPyV, as

for JCV and BKV. However, this hypothesis was not supported by a recent study

that reported KIPyV and WUPyV in autopsy lymphoid tissues of AIDS patients (13).

Whether WUPyV or KIPyV cause disease in HIV-positive persons or other

populations remains to be determined.

Our data demonstrate that WUPyV was detected in peripheral blood of HIV- and

HCV-infected persons. However, the scope of this study was limited because

clinical data were not available for study participants. Whether WUPyV or KIPyV

have oncogenicity or other pathogenicity in immunocompromised hosts remain to be

determined. The role of polyomaviruses in human cancers has been extensively

investigated but conclusive evidence is lacking (14). The genome of Merkel cell

polyomavirus, a new polyomavirus, was found to be integrated within the cellular

genome of Merkel cell carcinoma tissue samples, which suggests a role for this

virus in a specific tumor (15). Therefore, studies to assess the oncogenic

potential of WUPyV and KIPyV are also needed.

Acknowledgments

We thank for continued support, intellectual and scientific input,

and exchange and critical review of the data; and the staff of the Clinical

Virology Laboratory, Yale–New Haven Hospital, for assistance in this study.

This study was supported by National Institutes of Health grant T32 A107210-20

and a MedImmune pediatric fellowship grant.

Dr is a fellow in Pediatric Infectious Diseases at Yale University School

of Medicine. His research interests are the biology, epidemiology, and oncogenic

potential of human polyomaviruses.

References

Allander T, sson K, Gupta S, Bjerkner A, Gordana B, Perrson MA, et al.

Identification of a third human polyomavirus. J Virol. 2007;81:4130–6. PubMed

DOI

Gaynor AM, Nissen MD, Whiley DM, MacKay IM, Lambert SB, Wu G, et al.

Identification of a novel polyomavirus from patients with acute respiratory

tract infections. PLoS Pathog. 2007;3:e64. PubMed DOI

Norja P, Ubillos I, Templeton K, Simmonds P. No evidence for an association

between infections with WU and KI polyomaviruses and respiratory disease. J Clin

Virol. 2007;40:307–11. PubMed DOI

Le BM, Demertzis LM, Wu G, Tibbets RJ, Buller R, Arens MQ, et al. Clinical and

epidemiologic characterization of WU polyomavirus infection, St. Louis,

Missouri. Emerg Infect Dis. 2007;13:1936–8.

Bialasiewicz S, Whiley DM, Lambert SB, K, Bletchly C, Wang D, et al. A

newly reported human polyomavirus, KI virus, is present in the respiratory tract

of Australian children. J Clin Virol. 2007;40:15–8. PubMed DOI

Wattier RL, Vazquez MV, Weibel C, Shapiro ED, Ferguson D, Landry ML, et al. Role

of human polyomaviruses in respiratory tract disease in young children. Emerg

Infect Dis. 2008;14:1766–8. PubMed DOI

Khalili K, Gordon J, White MK. The polyomavirus, JCV and its involvement in

human disease. Adv Exp Med Biol. 2006;577:274–87. PubMed DOI

Hirsch HH. Polyomavirus BK nephropathy: a (re-)emerging complication in renal

transplantation. Am J Transplant. 2002;2:25–30. PubMed DOI

Andreoletti L, Lescieux A, Lambert B, Si-Mohamed A, Matta M, Wattre P, et al.

Semiquantitative detection of JCV-DNA in peripheral blood leukocytes from

HIV-1-infected patients with or without progressive multifocal

leukoencephalopathy. J Med Virol. 2002;66:1–7. PubMed DOI

Hymes LC, Warshaw BL. Polyomavirus (BK) in pediatric renal transplants:

evaluation of viremic patients with and without BK associated nephritis. Pediatr

Transplant. 2006;10:920–2. PubMed DOI

Stolt A, Sasnauskas K, Koskela P, Lehtinen M, Dillner J. Seroepidemiology of the

human polyomaviruses. J Gen Virol. 2003;84:1499–504. PubMed DOI

Costa C, Bergallo M, Astegiano S, Terlizzi ME, Sidoti F, Segoloni GP, et al.

Monitoring of BK virus replication in the first year following renal

transplantation. Nephrol Dial Transplant. 2008;23:3333–6. PubMed DOI

Sharp CP, Norja P, J, Bell JE, Simmonds P. Reactivation and mutation of

newly discovered WU, KI, and Merkel cell carcinoma polyomaviruses in

immunosuppressed individuals. J Infect Dis. 2009;199:398–404. PubMed DOI

zur Hausen H. Novel human polyomaviruses–re-emergence of a well known virus

family as possible human carcinogens. Int J Cancer. 2008;123:247–50. PubMed DOI

Feng H, Shuda M, Chang Y, PS. Clonal integration of a polyomavirus in

human Merkel cell carcinoma. Science. 2008;319:1096–100. PubMed DOI

Suggested Citation for this Article

MA, Weibel C, Ferguson D, Landry ML, Kahn JS. WU polyomavirus in patients

infected with HIV or hepatitis C virus, Connecticut, USA, 2007. Emerg Infect Dis

[serial on the Internet]. 2009 Jul [date cited]. Available from

http://www.cdc.gov/EID/content/15/7/1095.htm

DOI: 10.3201/eid1507.090150

S. Kahn, Department of Pediatrics, Division of Infectious Diseases, Yale

University School of Medicine, PO Box 208064, New Haven, CT 06520-8064, USA;

email: jeffrey.kahn@...

[Guest guest]

http://www.cdc.gov/eid/content/15/7/1095.htm

EID Journal Home> Volume 15, Number 7–July 2009

Volume 15, Number 7–July 2009

Dispatch

WU Polyomavirus in Patients Infected with HIV or Hepatitis C Virus, Connecticut,

USA, 2007

A. , Carla Weibel, Ferguson, Marie L. Landry, and S.

Kahn

Author affiliation: Yale University School of Medicine, New Haven, Connecticut,

USA

Abstract

WU polyomavirus (WUPyV) was detected in 10 (8.3%) of 121 HIV-positive plasma

specimens, 0 (0%) of 120 HIV-negative serum specimens, and 2 (2.5%) of 79

hepatitis C virus (HCV)–positive serum specimens. KI polyomavirus was not

detected in HIV-positive plasma or HCV-positive serum specimens. HIV-infected

persons may be susceptible to systemic WUPyV infection.

In 2007, 2 new human polyomaviruses, KI polyomavirus (KIPyV) and WU polyomavirus

(WUPyV), were identified. KIPyV was initially detected in an extract obtained

from 20 pooled randomly selected nasopharyngeal aspirates, and WUPyV was

detected in a nasopharyngeal aspirate from a 3-year-old child from Australia who

had a diagnosis of pneumonia (1,2). These viruses have since been detected in

respiratory tract specimens from symptomatic and asymptomatic children, although

no clear association with respiratory disease has been demonstrated (3–6).

Previously identified human polyomaviruses (BK virus [bKV] and JC virus [JCV])

cause clinical disease in immunocompromised persons (7,8). Although viremia may

be associated with immunosuppression, correlation of JCV DNA in peripheral blood

with development of progressive multifocal encephalopathy in AIDS patients

remains controversial (9). BKV DNA has been detected in blood of renal

transplant patients, and BKV load may be predictive of polyomavirus-associated

nephropathy (10).

The Study

The pathogenesis and clinical spectra of WUPyV and KIPyV, particularly in

immunocompromised persons, have not been defined. To investigate whether WUPyV

or KIPyV is present in persons with chronic viral infection and perhaps

compromised immunity, we conducted a cross-sectional study in which we screened

the following for WUPyV and KIPYV DNA: plasma samples from HIV-infected persons,

serum samples from hepatitis C virus (HCV)–infected persons, and a control group

of HIV-negative persons.

Three groups of samples submitted to the Clinical Virology Laboratory at

Yale–New Haven Hospital in 2007 were screened: HIV PCR-positive plasma, HCV

PCR-positive serum, and HIV antibody-negative serum. Patient identifiers were

removed and these specimens were tested as part of our ongoing investigation for

newly identified viruses. Collection of specimens and clinical data was approved

by the Yale University Human Investigation Committee and was compliant with

Health Insurance Portability and Accountability Act regulations.

Nucleic acids were extracted from each specimen by using QIAamp nucleic acid

purification kits (QIAGEN, Valencia, CA, USA). Screening for WUPyV DNA has been

described (6). Briefly, we performed an initial PCR screening specific for the

virus capsid protein 2 (VP2) gene by using primers described by Gaynor et al.

(2) and a nested PCR (Table 1, primers 1 and 2). To confirm results, DNA from

all PCR-positive samples was reextracted and screened with primers specific for

the region of the genome containing the noncoding control region (NCCR), which

includes the virus origin of replication (genome coordinates 5213 to nt 36 of

the circular viral genome). This screening included an initial PCR (Table 1,

primers 3 and 4) and a nested PCR (Table 1, primers 5 and 6). The nested PCR

generated a 328-bp amplicon. Screening for KIPyV DNA by PCR included a nested

PCR specific for the VP1 gene according to the protocol described by Allander et

al. (1). Positive and negative controls were included in each set of PCRs.

All PCR products were sequenced by using 377 DNA automated sequencers (Applied

Biosystems, City, CA, USA) at the W.M. Keck Biotechnology Resource

Laboratory at Yale University School of Medicine. For WUPyV, phylogenetic

analysis was performed on a 194-bp fragment within the amplified region of NCCR

(nt 5197 to nt 159 of the circular viral genome) by using Lasergene MegAlign

software (DNASTAR Inc., Madison, WI, USA) (ClustalW alignment method). The only

clinical data available for these deidentified serum specimens were HIV/HCV

status and virus loads. HIV and HCV virus loads were determined in the Clinical

Virology Laboratory by quantitative reverse transcription–PCR using commercially

available diagnostic tests. The Fisher exact test was used to determine whether

the difference in the percentage of WUPyV-positive specimens in HIV-positive and

HIV-negative patients was statistically significant.

Ten (8.3%) of 121 HIV-positive specimens and 0 (0%) of 120 HIV-negative samples

were positive for WUPyV (p85% of the persons screened will have detectable

antibodies to WUPyV (11). Whether antibody status for WUPyV correlates with

viremia is unknown. Viremia may represent primary infection or reactivation of

latent infection.

Viremia has been described for JCV and BKV. JCV DNA in serum/plasma may

correlate with the degree of immunosuppression. However, blood from viremic

persons infected with JCV has a low positive predictive value for development of

progressive multifocal encephalopathy in AIDS patients (9). A recent study

suggested that screening for BKV replication is useful in identifying patients

at risk for BKV-associated nephropathy, which may enable early interventions

such as renal biopsy and reduction of immunosuppression (12). However, because

of the lack of clinical data available for WUPyV-positive persons in our study,

it was not possible to make any clinical correlations.

The absence of KIPyV in HIV-positive or HCV-positive peripheral blood specimens

suggests that host susceptibility for KIPyV may differ from that of WUPyV, as

for JCV and BKV. However, this hypothesis was not supported by a recent study

that reported KIPyV and WUPyV in autopsy lymphoid tissues of AIDS patients (13).

Whether WUPyV or KIPyV cause disease in HIV-positive persons or other

populations remains to be determined.

Our data demonstrate that WUPyV was detected in peripheral blood of HIV- and

HCV-infected persons. However, the scope of this study was limited because

clinical data were not available for study participants. Whether WUPyV or KIPyV

have oncogenicity or other pathogenicity in immunocompromised hosts remain to be

determined. The role of polyomaviruses in human cancers has been extensively

investigated but conclusive evidence is lacking (14). The genome of Merkel cell

polyomavirus, a new polyomavirus, was found to be integrated within the cellular

genome of Merkel cell carcinoma tissue samples, which suggests a role for this

virus in a specific tumor (15). Therefore, studies to assess the oncogenic

potential of WUPyV and KIPyV are also needed.

Acknowledgments

We thank for continued support, intellectual and scientific input,

and exchange and critical review of the data; and the staff of the Clinical

Virology Laboratory, Yale–New Haven Hospital, for assistance in this study.

This study was supported by National Institutes of Health grant T32 A107210-20

and a MedImmune pediatric fellowship grant.

Dr is a fellow in Pediatric Infectious Diseases at Yale University School

of Medicine. His research interests are the biology, epidemiology, and oncogenic

potential of human polyomaviruses.

References

Allander T, sson K, Gupta S, Bjerkner A, Gordana B, Perrson MA, et al.

Identification of a third human polyomavirus. J Virol. 2007;81:4130–6. PubMed

DOI

Gaynor AM, Nissen MD, Whiley DM, MacKay IM, Lambert SB, Wu G, et al.

Identification of a novel polyomavirus from patients with acute respiratory

tract infections. PLoS Pathog. 2007;3:e64. PubMed DOI

Norja P, Ubillos I, Templeton K, Simmonds P. No evidence for an association

between infections with WU and KI polyomaviruses and respiratory disease. J Clin

Virol. 2007;40:307–11. PubMed DOI

Le BM, Demertzis LM, Wu G, Tibbets RJ, Buller R, Arens MQ, et al. Clinical and

epidemiologic characterization of WU polyomavirus infection, St. Louis,

Missouri. Emerg Infect Dis. 2007;13:1936–8.

Bialasiewicz S, Whiley DM, Lambert SB, K, Bletchly C, Wang D, et al. A

newly reported human polyomavirus, KI virus, is present in the respiratory tract

of Australian children. J Clin Virol. 2007;40:15–8. PubMed DOI

Wattier RL, Vazquez MV, Weibel C, Shapiro ED, Ferguson D, Landry ML, et al. Role

of human polyomaviruses in respiratory tract disease in young children. Emerg

Infect Dis. 2008;14:1766–8. PubMed DOI

Khalili K, Gordon J, White MK. The polyomavirus, JCV and its involvement in

human disease. Adv Exp Med Biol. 2006;577:274–87. PubMed DOI

Hirsch HH. Polyomavirus BK nephropathy: a (re-)emerging complication in renal

transplantation. Am J Transplant. 2002;2:25–30. PubMed DOI

Andreoletti L, Lescieux A, Lambert B, Si-Mohamed A, Matta M, Wattre P, et al.

Semiquantitative detection of JCV-DNA in peripheral blood leukocytes from

HIV-1-infected patients with or without progressive multifocal

leukoencephalopathy. J Med Virol. 2002;66:1–7. PubMed DOI

Hymes LC, Warshaw BL. Polyomavirus (BK) in pediatric renal transplants:

evaluation of viremic patients with and without BK associated nephritis. Pediatr

Transplant. 2006;10:920–2. PubMed DOI

Stolt A, Sasnauskas K, Koskela P, Lehtinen M, Dillner J. Seroepidemiology of the

human polyomaviruses. J Gen Virol. 2003;84:1499–504. PubMed DOI

Costa C, Bergallo M, Astegiano S, Terlizzi ME, Sidoti F, Segoloni GP, et al.

Monitoring of BK virus replication in the first year following renal

transplantation. Nephrol Dial Transplant. 2008;23:3333–6. PubMed DOI

Sharp CP, Norja P, J, Bell JE, Simmonds P. Reactivation and mutation of

newly discovered WU, KI, and Merkel cell carcinoma polyomaviruses in

immunosuppressed individuals. J Infect Dis. 2009;199:398–404. PubMed DOI

zur Hausen H. Novel human polyomaviruses–re-emergence of a well known virus

family as possible human carcinogens. Int J Cancer. 2008;123:247–50. PubMed DOI

Feng H, Shuda M, Chang Y, PS. Clonal integration of a polyomavirus in

human Merkel cell carcinoma. Science. 2008;319:1096–100. PubMed DOI

Suggested Citation for this Article

MA, Weibel C, Ferguson D, Landry ML, Kahn JS. WU polyomavirus in patients

infected with HIV or hepatitis C virus, Connecticut, USA, 2007. Emerg Infect Dis

[serial on the Internet]. 2009 Jul [date cited]. Available from

http://www.cdc.gov/EID/content/15/7/1095.htm

DOI: 10.3201/eid1507.090150

S. Kahn, Department of Pediatrics, Division of Infectious Diseases, Yale

University School of Medicine, PO Box 208064, New Haven, CT 06520-8064, USA;

email: jeffrey.kahn@...

[Guest guest]

http://www.cdc.gov/eid/content/15/7/1095.htm

EID Journal Home> Volume 15, Number 7–July 2009

Volume 15, Number 7–July 2009

Dispatch

WU Polyomavirus in Patients Infected with HIV or Hepatitis C Virus, Connecticut,

USA, 2007

A. , Carla Weibel, Ferguson, Marie L. Landry, and S.

Kahn

Author affiliation: Yale University School of Medicine, New Haven, Connecticut,

USA

Abstract

WU polyomavirus (WUPyV) was detected in 10 (8.3%) of 121 HIV-positive plasma

specimens, 0 (0%) of 120 HIV-negative serum specimens, and 2 (2.5%) of 79

hepatitis C virus (HCV)–positive serum specimens. KI polyomavirus was not

detected in HIV-positive plasma or HCV-positive serum specimens. HIV-infected

persons may be susceptible to systemic WUPyV infection.

In 2007, 2 new human polyomaviruses, KI polyomavirus (KIPyV) and WU polyomavirus

(WUPyV), were identified. KIPyV was initially detected in an extract obtained

from 20 pooled randomly selected nasopharyngeal aspirates, and WUPyV was

detected in a nasopharyngeal aspirate from a 3-year-old child from Australia who

had a diagnosis of pneumonia (1,2). These viruses have since been detected in

respiratory tract specimens from symptomatic and asymptomatic children, although

no clear association with respiratory disease has been demonstrated (3–6).

Previously identified human polyomaviruses (BK virus [bKV] and JC virus [JCV])

cause clinical disease in immunocompromised persons (7,8). Although viremia may

be associated with immunosuppression, correlation of JCV DNA in peripheral blood

with development of progressive multifocal encephalopathy in AIDS patients

remains controversial (9). BKV DNA has been detected in blood of renal

transplant patients, and BKV load may be predictive of polyomavirus-associated

nephropathy (10).

The Study

The pathogenesis and clinical spectra of WUPyV and KIPyV, particularly in

immunocompromised persons, have not been defined. To investigate whether WUPyV

or KIPyV is present in persons with chronic viral infection and perhaps

compromised immunity, we conducted a cross-sectional study in which we screened

the following for WUPyV and KIPYV DNA: plasma samples from HIV-infected persons,

serum samples from hepatitis C virus (HCV)–infected persons, and a control group

of HIV-negative persons.

Three groups of samples submitted to the Clinical Virology Laboratory at

Yale–New Haven Hospital in 2007 were screened: HIV PCR-positive plasma, HCV

PCR-positive serum, and HIV antibody-negative serum. Patient identifiers were

removed and these specimens were tested as part of our ongoing investigation for

newly identified viruses. Collection of specimens and clinical data was approved

by the Yale University Human Investigation Committee and was compliant with

Health Insurance Portability and Accountability Act regulations.

Nucleic acids were extracted from each specimen by using QIAamp nucleic acid

purification kits (QIAGEN, Valencia, CA, USA). Screening for WUPyV DNA has been

described (6). Briefly, we performed an initial PCR screening specific for the

virus capsid protein 2 (VP2) gene by using primers described by Gaynor et al.

(2) and a nested PCR (Table 1, primers 1 and 2). To confirm results, DNA from

all PCR-positive samples was reextracted and screened with primers specific for

the region of the genome containing the noncoding control region (NCCR), which

includes the virus origin of replication (genome coordinates 5213 to nt 36 of

the circular viral genome). This screening included an initial PCR (Table 1,

primers 3 and 4) and a nested PCR (Table 1, primers 5 and 6). The nested PCR

generated a 328-bp amplicon. Screening for KIPyV DNA by PCR included a nested

PCR specific for the VP1 gene according to the protocol described by Allander et

al. (1). Positive and negative controls were included in each set of PCRs.

All PCR products were sequenced by using 377 DNA automated sequencers (Applied

Biosystems, City, CA, USA) at the W.M. Keck Biotechnology Resource

Laboratory at Yale University School of Medicine. For WUPyV, phylogenetic

analysis was performed on a 194-bp fragment within the amplified region of NCCR

(nt 5197 to nt 159 of the circular viral genome) by using Lasergene MegAlign

software (DNASTAR Inc., Madison, WI, USA) (ClustalW alignment method). The only

clinical data available for these deidentified serum specimens were HIV/HCV

status and virus loads. HIV and HCV virus loads were determined in the Clinical

Virology Laboratory by quantitative reverse transcription–PCR using commercially

available diagnostic tests. The Fisher exact test was used to determine whether

the difference in the percentage of WUPyV-positive specimens in HIV-positive and

HIV-negative patients was statistically significant.

Ten (8.3%) of 121 HIV-positive specimens and 0 (0%) of 120 HIV-negative samples

were positive for WUPyV (p85% of the persons screened will have detectable

antibodies to WUPyV (11). Whether antibody status for WUPyV correlates with

viremia is unknown. Viremia may represent primary infection or reactivation of

latent infection.

Viremia has been described for JCV and BKV. JCV DNA in serum/plasma may

correlate with the degree of immunosuppression. However, blood from viremic

persons infected with JCV has a low positive predictive value for development of

progressive multifocal encephalopathy in AIDS patients (9). A recent study

suggested that screening for BKV replication is useful in identifying patients

at risk for BKV-associated nephropathy, which may enable early interventions

such as renal biopsy and reduction of immunosuppression (12). However, because

of the lack of clinical data available for WUPyV-positive persons in our study,

it was not possible to make any clinical correlations.

The absence of KIPyV in HIV-positive or HCV-positive peripheral blood specimens

suggests that host susceptibility for KIPyV may differ from that of WUPyV, as

for JCV and BKV. However, this hypothesis was not supported by a recent study

that reported KIPyV and WUPyV in autopsy lymphoid tissues of AIDS patients (13).

Whether WUPyV or KIPyV cause disease in HIV-positive persons or other

populations remains to be determined.

Our data demonstrate that WUPyV was detected in peripheral blood of HIV- and

HCV-infected persons. However, the scope of this study was limited because

clinical data were not available for study participants. Whether WUPyV or KIPyV

have oncogenicity or other pathogenicity in immunocompromised hosts remain to be

determined. The role of polyomaviruses in human cancers has been extensively

investigated but conclusive evidence is lacking (14). The genome of Merkel cell

polyomavirus, a new polyomavirus, was found to be integrated within the cellular

genome of Merkel cell carcinoma tissue samples, which suggests a role for this

virus in a specific tumor (15). Therefore, studies to assess the oncogenic

potential of WUPyV and KIPyV are also needed.

Acknowledgments

We thank for continued support, intellectual and scientific input,

and exchange and critical review of the data; and the staff of the Clinical

Virology Laboratory, Yale–New Haven Hospital, for assistance in this study.

This study was supported by National Institutes of Health grant T32 A107210-20

and a MedImmune pediatric fellowship grant.

Dr is a fellow in Pediatric Infectious Diseases at Yale University School

of Medicine. His research interests are the biology, epidemiology, and oncogenic

potential of human polyomaviruses.

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Identification of a novel polyomavirus from patients with acute respiratory

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Norja P, Ubillos I, Templeton K, Simmonds P. No evidence for an association

between infections with WU and KI polyomaviruses and respiratory disease. J Clin

Virol. 2007;40:307–11. PubMed DOI

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epidemiologic characterization of WU polyomavirus infection, St. Louis,

Missouri. Emerg Infect Dis. 2007;13:1936–8.

Bialasiewicz S, Whiley DM, Lambert SB, K, Bletchly C, Wang D, et al. A

newly reported human polyomavirus, KI virus, is present in the respiratory tract

of Australian children. J Clin Virol. 2007;40:15–8. PubMed DOI

Wattier RL, Vazquez MV, Weibel C, Shapiro ED, Ferguson D, Landry ML, et al. Role

of human polyomaviruses in respiratory tract disease in young children. Emerg

Infect Dis. 2008;14:1766–8. PubMed DOI

Khalili K, Gordon J, White MK. The polyomavirus, JCV and its involvement in

human disease. Adv Exp Med Biol. 2006;577:274–87. PubMed DOI

Hirsch HH. Polyomavirus BK nephropathy: a (re-)emerging complication in renal

transplantation. Am J Transplant. 2002;2:25–30. PubMed DOI

Andreoletti L, Lescieux A, Lambert B, Si-Mohamed A, Matta M, Wattre P, et al.

Semiquantitative detection of JCV-DNA in peripheral blood leukocytes from

HIV-1-infected patients with or without progressive multifocal

leukoencephalopathy. J Med Virol. 2002;66:1–7. PubMed DOI

Hymes LC, Warshaw BL. Polyomavirus (BK) in pediatric renal transplants:

evaluation of viremic patients with and without BK associated nephritis. Pediatr

Transplant. 2006;10:920–2. PubMed DOI

Stolt A, Sasnauskas K, Koskela P, Lehtinen M, Dillner J. Seroepidemiology of the

human polyomaviruses. J Gen Virol. 2003;84:1499–504. PubMed DOI

Costa C, Bergallo M, Astegiano S, Terlizzi ME, Sidoti F, Segoloni GP, et al.

Monitoring of BK virus replication in the first year following renal

transplantation. Nephrol Dial Transplant. 2008;23:3333–6. PubMed DOI

Sharp CP, Norja P, J, Bell JE, Simmonds P. Reactivation and mutation of

newly discovered WU, KI, and Merkel cell carcinoma polyomaviruses in

immunosuppressed individuals. J Infect Dis. 2009;199:398–404. PubMed DOI

zur Hausen H. Novel human polyomaviruses–re-emergence of a well known virus

family as possible human carcinogens. Int J Cancer. 2008;123:247–50. PubMed DOI

Feng H, Shuda M, Chang Y, PS. Clonal integration of a polyomavirus in

human Merkel cell carcinoma. Science. 2008;319:1096–100. PubMed DOI

Suggested Citation for this Article

MA, Weibel C, Ferguson D, Landry ML, Kahn JS. WU polyomavirus in patients

infected with HIV or hepatitis C virus, Connecticut, USA, 2007. Emerg Infect Dis

[serial on the Internet]. 2009 Jul [date cited]. Available from

http://www.cdc.gov/EID/content/15/7/1095.htm

DOI: 10.3201/eid1507.090150

S. Kahn, Department of Pediatrics, Division of Infectious Diseases, Yale

University School of Medicine, PO Box 208064, New Haven, CT 06520-8064, USA;

email: jeffrey.kahn@...