PBMC

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What kind of Class I CEF-peptides are you testing for?

Category: PBMC

Can these cells (PBMC) be cultured and for how long? What is the growth rate? How do you culture them?

Yes, it should be easily possible to generate and grow antigen specific memory cells

First the cells need to be stimulated with an antigen is assay conditions (CTL recommends 300,000 cells per ml stimulated with peptides around 2µg/ml concentration. After 4 days cells need to be counted and viable cells be plated 3x105/ml and biological grade active IL-2 to be given at 2units/ml. Follow the cell lines growth for 5 days by looking at them with the microscope regularly Again count spin down and provide fresh IL-2 medium 3x105/ml Follow the cell lines growth for an additional 5 days and if they still look good and the cell number increases If the cell number start to decline cells need to be stimulated with antigen and irradiated autologus PBMC, this time without adding IL-2 into the medium and start from the beginning.

The T cell lines need to be restimulated with the antigen and the Peptide in the presence of irradiated autologus PBMC.

Literature:

Tary-Lehmann, M., Rolink, A.G., Lehmann, P.V., Nagy, Z.A., and Hurtenbach, U. Induction of graft versus host-associated immunodeficiency by CD4+ T-cell clones.  J. Immunol.  1990.  145:2092-2098.

Rininsland F.H., Helms T., Asaad R.J., Boehm B.O. and Tary-Lehmann M.  Granzyme B ELISPOT assay for ex vivo measurement of T cell immunity.  J. Immunol. Methods 240:143-155, 2000.

Category: PBMC

What kind of HLA-typing results do you provide?

Samples have been HLA-typed low and high resolution results are available for 3 Class I loci (A, B, C) and 4 Class II loci (DRB1, DQA, DQB and DP)

Category: PBMC

What kind of protein-antigens are you testing for?

6 protein-antigens: Candida, PPD, Mosquito, Dust Mite, Mumps, PHA

Category: PBMC

What kind of peptide pools are you testing for?

Two Class I CEF-Peptide Pools (23 and 32) and one Class II CEF-Peptide Pool.

Category: PBMC

What kinds of flu strains are used in immune characterization?

Category: PBMC

The responses you have seen with the above antigens are for which cytokines?

IFN-gamma

Category: PBMC

What kind of anticoagulant has been used?

ACDA present in GAMBRO bags for blood collections.

Category: PBMC

What kind of machine has been used for collection by aphaeresis?

COP Spectra machine

Category: PBMC

Are the PBMC samples tested for presence of EBV – sequences by PCR?

No, they are not.

Category: PBMC

Is there a serological profile of each donor; are the donors EBV-seronegative?

No, we don’t have serological profiles.

Category: PBMC

Can you guarantee continuous delivery of PBMC over a long time span?

Yes, we can deliver virtually any number of vials any time. We can also freeze the requested cells and deliver them within a customer-specified time range and release dates. Remember that CTL's proprietary freezing protocols ensure viability rates that are comparable to those of fresh PBMC samples.

Category: PBMC

How do you ship these cells?

CTL is using Dry Cryoshippers for shipping and ships them overnight with next morning delivery. These Cryoshippers are being used because the shipping cost is significantly lower then shipping them on Dry Ice and the shipping conditions are the same as storage conditions, therefore the character of the cells stays the same.

Category: PBMC

What is the viability rate of the cells after thawing?

Frozen PBMC maintain >90% viability after thawing when CTL-Thawing Protocol is followed (see attached info: “Fresh vs. Frozen” poster)

Category: PBMC

What kind of laboratory testings has been done for the LP donors?

For each donor in our PBMC database, the following tests have been performed:

          ABO and RH Group/Type

          HIV 1/2 Antibody

          Irregular Antibody Screening

          HTLV I/II Antibody

          Hepatitis B Surface Antigen

          HCV Antibody

          Hepatitis B Core Antibody

          Serologic Test for Syphilis

          NAT HIV/HCV

Category: PBMC