Another debate in viral vaccines: nanobacterial contamination


http://www.asmusa.org/pcsrc/gm2001/33749.htm

101st General Meeting of the American Society for Microbiology

May 20-24, 2001, Orlando, Florida

For more information on any presentation at the 101st General Meeting conta=
ct Jim Sliwa, ASM Communications at jsliwa@asmusa.org.


(Session 078/Y , Paper Y-3)

Neva Ciftcioglu
University of Kuopio
Kuopio,
Finland
35817163054
neva.ciftcioglu@uku.fi

Vaccines are an indispensable weapon in the battle to prevent disease, both=
in humans and animals. Provision and protection of the vaccine supply is so=
essential that no possible pathogen is beyond suspicion. So it is with nano=
bacteria (NB), a potential infectious agent so new that science is still deb=
ating its existence as a "living organism". We analysed 6 veterinary vaccine=
s and 3 inactivated human polio vaccines produced in cell culture for NB. We=
report that 3 of 6 European veterinary vaccines contained NB. Of the 3 dist=
inct lots of polio vaccine from European manufacturers, 2 lots from manufact=
urer-1 were NB-positive and 1 lot from manufacturer-2 was NB-negative. These=
results suggest that not all lots of vaccine contain detectable NB. The pub=
lic health risk, if any, from nanobacteria is yet to be defined, but nanobac=
teria have been found in kidney stones, liver and kidney cyst fluids, and im=
plicated in kidney stone formation.

In the early 1990's, Drs. Olavi Kajander and Neva Ciftcioglu, University of=
Kuopio, Finland discovered a minute self replicating agent, which was named=
"nanobacteria", in fetal bovine serum (FBS) that mediates mineral (calcium =
phosphate) formation under conditions found in blood and urine. NB was subse=
quently found to contaminate cell cultures widely used in research and to pa=
ss through filters commonly used to sterilize vaccines. Several vaccines are=
produced in mammalian cell cultures that were initially or are currently cu=
ltured in media supplemented with FBS. Thus, the question arose, "Could the =
NB found in some batches of FBS contaminate some vaccines"?

To detect nanobacteria in these 9 vaccines, the Kuopio University team cult=
ured the vaccines in routine tissue culture medium known by prior testing to=
be free of NB and examined the original vaccines and cultured vaccines by e=
lectron microscopy, and immunologic tests for NB antigens (immunofluorescent=
staining, dot blot assay and ELISA). NB were found 3 of 6 vet vaccine produ=
cts and 2 of 3 human vaccine products prepared using FBS. A second question =
arose, "Even if NB are present in the culture broth, why were they not remov=
ed during the purification and sterilizing steps of vaccine production"?

Cell culture mediated viral vaccine production requires the use of cells to=
propagate large amounts of virus, which is then filtered to remove contamin=
ates and cells present in the propagation step. The viruses passing through =
the filters are then further processed and inactivated prior to use. NB are =
in the same size range as certain large viruses, and may adhere to viruses t=
hus escaping purification steps using filtration. NB are also resistant to m=
any disinfecting chemicals and antibiotics. NB can exist intracellularly and=
grow in the absence of FBS. At this time it is not known if FBS added to th=
e production cell lines or NB in the cells lines themselves are the source o=
f NB contamination in these cases.

The Center for Biologics Evaluation and Research (CBER) and the U.S. Food a=
nd Drug Administration (FDA) learned earlier this year that some vaccines we=
re manufactured with bovine–derived materials obtained from countries where =
bovine spongiform encephalopathy (BSE; mad cow disease) is prevalent or wher=
e a substantial risk for BSE exists. Creutzfeldt-Jakob disease (vCJD), the h=
uman equivalent of BSE, has been attributed to, among other possibilities, e=
ating beef products from cattle infected with the prion agent of BSE. Althou=
gh there is no evidence to date that cases of vCJD are related to the use of=
vaccines, interestingly a vaccine lot was withdrawn because of being expose=
d to FBS from the country where high risk for this prion disease exists.

The study summarised in the first paragraph will be presented in the 101st =
ASM Meeting in Orlando in May 21, 2001 (1.00-2.30pm). The Kuopio team's mess=
age to the scientific society in this presentation will be: More research is=
needed to understand nanobacteria and their role in human and animal diseas=
es. To err on the side of safety is no vice; just as prions and viruses were=
viewed with scepticism before they were completely characterised, so concep=
ts of nanobacteria (and the scientists who study them) will be knocked aroun=
d until more is known about NB. Vaccines should be regarded as safe and esse=
ntial to human health in the modern age until they are proven otherwise.

Related Literature:

NB have been linked to kidney stones (Ciftcioglu et al., Kidney Internation=
al 56:1893-8,1999), as a kidney stone causative agent (Garcia Cuerpo et al.,=
Arch Esp Urol 53:291-303, 2000), and in polycystic kidney disease (Hjelle e=
t al, Kidney International 57: 2360-2374, 2000).

For a recent review of nanobacteria, (Kajander, Ciftcioglu, Miller-Hjelle, =
Hjelle. Current Opinions in Nephrology and Hypertension 2001 in press).