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Dendritic cell vaccines containing lymphocytes produce improved immunogenicity in patients with cancer.

Frank MO, Kaufman J, Parveen S, Blachère NE, Orange DE, Darnell RB - J Transl Med (2014)

Bottom Line: Dendritic cells are currently under investigation for their ability to generate anti-cancer immune responses.No consensus has been reached as to the optimal method of dendritic cell vaccine preparation and is a barrier to success in the field.This difference could not be accounted for by dendritic cell vaccine dose, cell surface phenotype or dendritic cell function in vitro.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Neuro-oncology, The Rockefeller University, 1230 York Avenue, New York, NY, 10065, USA. frankm@rockefeller.edu.

ABSTRACT

Background: Dendritic cells are currently under investigation for their ability to generate anti-cancer immune responses. No consensus has been reached as to the optimal method of dendritic cell vaccine preparation and is a barrier to success in the field.

Methods: Over a course of three separate dendritic cell vaccine studies to treat cancer, we tested two different methods for preparing dendritic cells from peripheral blood mononuclear cells: adherence and antibody-selected CD14+ cells.

Results: Surprisingly, we found that patients who received dendritic cell vaccines generated by the adherence method mounted increased T cell proliferation in response to vaccination. This difference could not be accounted for by dendritic cell vaccine dose, cell surface phenotype or dendritic cell function in vitro. One notable difference between the two vaccine preparation methods was that the dendritic cell vaccine cultures generated by the adherence method contained up to 10% lymphocytes, and these lymphocytes were proliferating and producing IFNγ in response to antigen in vitro at the time of administration.

Conclusions: Enhanced immunogenicity of adherence dendritic cell vaccinations may be due to the presence of lymphocytes during dendritic cell culture.

Trial registration: Clinicaltrials.gov identifiers: NCT00289341, NCT00345293, and NCT00893945.

No MeSH data available.


Related in: MedlinePlus

DC vaccines prepared by Adherence support lymphocyte proliferation and cytokine production. (A) DCs made by the 2 methods were assessed for proliferation by 3H thymidine incorporation at 48, 72, and 96 hours beyond day 8, (the day DCs would have been administered as vaccine). Proliferation responses for Adherence DCs are shown in red and Selected DCs are shown in purple. The results are averages of triplicate wells. * indicates p < 0.05. (B) Supernatants of Adherence and Selected DC cultures were examined for pro-inflammatory cytokines on Day 6 and Day 8. Data for IL-2 is shown for 3 donors. Black line represents the mean. * indicates p < 0.05. NS = not statistically significant.
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Fig4: DC vaccines prepared by Adherence support lymphocyte proliferation and cytokine production. (A) DCs made by the 2 methods were assessed for proliferation by 3H thymidine incorporation at 48, 72, and 96 hours beyond day 8, (the day DCs would have been administered as vaccine). Proliferation responses for Adherence DCs are shown in red and Selected DCs are shown in purple. The results are averages of triplicate wells. * indicates p < 0.05. (B) Supernatants of Adherence and Selected DC cultures were examined for pro-inflammatory cytokines on Day 6 and Day 8. Data for IL-2 is shown for 3 donors. Black line represents the mean. * indicates p < 0.05. NS = not statistically significant.

Mentions: To assess lymphocyte proliferation within the DC preparations, both Adherence and Selected DCs were pulsed with 3H-thymidine. Adherence DC preparations were found to have incorporated more 3H-thymidine than Selected DCs, an indication that the lymphocytes were proliferating in these cultures (Figure 4A, p = 0.03, p = 0.003, and p = 0.021 respectively at 48, 72, and 96 hours). In addition, when we examined the Adherence and Selected DC cultures for the presence of cytokines after 6 days in culture, we found that the Adherence DC supernatants contained a mean of 4.42 pg/ml IL-2 while the levels in the Selected DC culture supernatants were below the level of detection (<0.68 pg/ml), a statistically significantly increase (Figure 4B, p = 0.012). This supports the observation that lymphocytes present in the Adherence DC culture were proliferating. There was a similar trend after 2 more days of culture in fresh media but this difference was not statistically significant (p = 0.070). There were no differences in the concentrations of IL-8, IL-12p70, IL-1β, IFNγ, IL-6, and IL-10 found in the supernatants of Adherence or Selected DC cultures. In conclusion, Adherence DC vaccines contain both DCs as well as CD4+, CD8+ and CD19+ lymphocytes that are activated in vitro, are proliferating, and are producing IL-2.Figure 4


Dendritic cell vaccines containing lymphocytes produce improved immunogenicity in patients with cancer.

Frank MO, Kaufman J, Parveen S, Blachère NE, Orange DE, Darnell RB - J Transl Med (2014)

DC vaccines prepared by Adherence support lymphocyte proliferation and cytokine production. (A) DCs made by the 2 methods were assessed for proliferation by 3H thymidine incorporation at 48, 72, and 96 hours beyond day 8, (the day DCs would have been administered as vaccine). Proliferation responses for Adherence DCs are shown in red and Selected DCs are shown in purple. The results are averages of triplicate wells. * indicates p < 0.05. (B) Supernatants of Adherence and Selected DC cultures were examined for pro-inflammatory cytokines on Day 6 and Day 8. Data for IL-2 is shown for 3 donors. Black line represents the mean. * indicates p < 0.05. NS = not statistically significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4264264&req=5

Fig4: DC vaccines prepared by Adherence support lymphocyte proliferation and cytokine production. (A) DCs made by the 2 methods were assessed for proliferation by 3H thymidine incorporation at 48, 72, and 96 hours beyond day 8, (the day DCs would have been administered as vaccine). Proliferation responses for Adherence DCs are shown in red and Selected DCs are shown in purple. The results are averages of triplicate wells. * indicates p < 0.05. (B) Supernatants of Adherence and Selected DC cultures were examined for pro-inflammatory cytokines on Day 6 and Day 8. Data for IL-2 is shown for 3 donors. Black line represents the mean. * indicates p < 0.05. NS = not statistically significant.
Mentions: To assess lymphocyte proliferation within the DC preparations, both Adherence and Selected DCs were pulsed with 3H-thymidine. Adherence DC preparations were found to have incorporated more 3H-thymidine than Selected DCs, an indication that the lymphocytes were proliferating in these cultures (Figure 4A, p = 0.03, p = 0.003, and p = 0.021 respectively at 48, 72, and 96 hours). In addition, when we examined the Adherence and Selected DC cultures for the presence of cytokines after 6 days in culture, we found that the Adherence DC supernatants contained a mean of 4.42 pg/ml IL-2 while the levels in the Selected DC culture supernatants were below the level of detection (<0.68 pg/ml), a statistically significantly increase (Figure 4B, p = 0.012). This supports the observation that lymphocytes present in the Adherence DC culture were proliferating. There was a similar trend after 2 more days of culture in fresh media but this difference was not statistically significant (p = 0.070). There were no differences in the concentrations of IL-8, IL-12p70, IL-1β, IFNγ, IL-6, and IL-10 found in the supernatants of Adherence or Selected DC cultures. In conclusion, Adherence DC vaccines contain both DCs as well as CD4+, CD8+ and CD19+ lymphocytes that are activated in vitro, are proliferating, and are producing IL-2.Figure 4

Bottom Line: Dendritic cells are currently under investigation for their ability to generate anti-cancer immune responses.No consensus has been reached as to the optimal method of dendritic cell vaccine preparation and is a barrier to success in the field.This difference could not be accounted for by dendritic cell vaccine dose, cell surface phenotype or dendritic cell function in vitro.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Neuro-oncology, The Rockefeller University, 1230 York Avenue, New York, NY, 10065, USA. frankm@rockefeller.edu.

ABSTRACT

Background: Dendritic cells are currently under investigation for their ability to generate anti-cancer immune responses. No consensus has been reached as to the optimal method of dendritic cell vaccine preparation and is a barrier to success in the field.

Methods: Over a course of three separate dendritic cell vaccine studies to treat cancer, we tested two different methods for preparing dendritic cells from peripheral blood mononuclear cells: adherence and antibody-selected CD14+ cells.

Results: Surprisingly, we found that patients who received dendritic cell vaccines generated by the adherence method mounted increased T cell proliferation in response to vaccination. This difference could not be accounted for by dendritic cell vaccine dose, cell surface phenotype or dendritic cell function in vitro. One notable difference between the two vaccine preparation methods was that the dendritic cell vaccine cultures generated by the adherence method contained up to 10% lymphocytes, and these lymphocytes were proliferating and producing IFNγ in response to antigen in vitro at the time of administration.

Conclusions: Enhanced immunogenicity of adherence dendritic cell vaccinations may be due to the presence of lymphocytes during dendritic cell culture.

Trial registration: Clinicaltrials.gov identifiers: NCT00289341, NCT00345293, and NCT00893945.

No MeSH data available.


Related in: MedlinePlus