Table 1.
Characteristics and use of PBMC as a study model for T. gondii and other Apicomplexan parasites during the last 10 years.
| Organism | Technical observations | PBMC culture (number of cells/final volume/culture plates) | Main findings | References |
|---|---|---|---|---|
| Human PBMC and Toxoplasma gondii |
Stimulus: RH strain (TLA, 1 μg/mL) Technic: RT-qPCR Process time: 48 h after collection Cryopreserved: No Supplemented: 10% FCS |
1 × 106/500 μL/48-well | High levels of TGF-β, IL-6, IL-10 in OT individuals | Maia et al., 2017 |
|
Stimulus: RH strain (TLA, 1 μg/mL) Technic: RT-qPCR Process time: 48 h after collection Cryopreserved: No Supplemented: 10% FBS |
1 × 106/500 μL/48-well | TATA box-binding protein (TBP) and ubiquitin C (UBC) are the most stable genes for mRNA analysis in PBMCs | Meira-Strejevitch et al., 2017 | |
|
Stimulus: ND Technic: Radioactivity Process time: 168 h in incubation Cryopreserved: No Supplemented: 5% human AB serum |
ND | Despite high proliferation, lymphocytes from meth users had a lower proliferative capacity | Massanella et al., 2015 | |
|
Stimulus: Peptides from P30 and ROP 18 (10 μg/mL) Technic: ELISPOT Process time: 24 h in incubation Cryopreserved: Yes Supplemented: No |
2 × 105/100 μL/96-well | Four peptides induced IFN-γ expression | Cardona et al., 2015 | |
|
Stimulus: RH strain (TLA, 1 μg/mL) Technic: ELISA Process time: 48 h in incubation Cryopreserved: No Supplemented: 10% FBS |
1 × 106/500 μL/48-well | Restoration of IFN-γ response and a decrease of the inflammatory cytokines TNF-α and IL-10 | Meira et al., 2015 | |
|
Stimulus: RH strain (Live) Technic: Radioactivity Process time: ND Cryopreserved: ND Supplemented: ND |
2.5 × 105/100 μL/96-well | Phytoecdysteroids did not inhibit Toxoplasma and did not affect the cytokine response (IFN-γ. IL-12, IL-10) | Dzitko et al., 2015 | |
|
Stimulus: TLA (ND) Technic: ELISPOT Process time: 120 h in incubation Cryopreserved: No Supplemented: No |
3 × 105/200 μL/ND | No association was observed when PBMCs were stimulated with TLA or mitogen | Nogueira et al., 2014 | |
|
Stimulus: BK strain (Live) Technic: ELISA Process time: 48 h in incubation Cryopreserved: No Supplemented: 0.1% BSA |
2.5 × 106/ND/ND | Correlation between Prolactine and the level of IL-10, but not with IFN-γ | Dzitko et al., 2012 | |
|
Stimulus: RH and ME49 strains (Live) Technic: ELISA Process time: 48 h in incubation Cryopreserved: No Supplemented: No |
2 × 106/ND/24-well | T. gondii-seronegative non-pregnant women produced significantly higher levels of TNF-a and IL-12 | Rezende-Oliveira et al., 2012 | |
|
Stimulus:
T. gondii peptides Technic: ELISPOT Process time: ND Cryopreserved: Yes Supplemented: No |
2 × 105/100 μL/96-well | Peptides induced significant IFN-γ production by PBMCs from 4 HLA-A*0201 persons infected with T. gondii | Cong et al., 2011 | |
|
Stimulus: TRRH strain (TLA, 5 mg/mL) Technic: ELISA Process time: 72 h in incubation Cryopreserved: No Supplemented: 10% FBS |
1 × 106/200 μL/96-well | IL-5 was higher than IFN-γ in the initial phase of the infection; as the IgG started to rise, IFN-γ increased and suppressed the synthesis of IL-5 | Bayram Delibaş et al., 2009 | |
| Pigs PBMC and Toxoplasma gondii |
Stimulus: IPB-G/LR strain (TLA, 10 μg/mL) Technic: Flow cytometry Process time: 72 h in incubation Cryopreserved: No Supplemented: 10% FBS |
1 × 106/ND/ND | High levels of IFN-γ | Jennes et al., 2017 |
|
Stimulus: RH strain (TLA) Technic: RNAseq Process time: 8, 24, 48 h in incubation Cryopreserved: No Supplemented: 10% FBS |
ND | More than 2,400 differentially expressed genes | Zhou et al., 2016a | |
| Human PBMC and Plasmodium sp. |
Stimulus:
P. falciparum crude lysate (50 mg) Technic: Flow cytometry and RT-qPCR Process time: 168 h in incubation Cryopreserved: No Supplemented: 10% FCS |
1 × 106/ND/ND | Crude antigens exhibited strong heterogeneity in the cytokine production | Kijogi et al., 2018 |
|
Stimulus:
P. falciparum crude lysate (50 mg) Technic: Flow cytometry and RT-qPCR Process time: 10 h in incubation Cryopreserved: No Supplemented: 2% FCS |
ND | Decreased parasite growth and expression of PD-1 and IL-10 genes using L-citrulline supplemented media | Awasthi et al., 2017 | |
|
Stimulus: Peptide Pooling Scheme Technic: ELISPOT Process time: 18 h in incubation Cryopreserved: ND Supplemented: 10% FCS |
1 × 106/ND/96-well | Highest immunogenicity was identified at 7 days after boosting with 932 SFC compared with 57 SFC among control vaccinees | Mensah et al., 2016 | |
|
Stimulus: Recombinant RAMA protein Technic: ELISA Process time: ND Cryopreserved: ND Supplemented: ND |
ND | High levels of interferon (IFN)-γ and interleukin (IL)-10 cytokines were detected | Changrob et al., 2016 | |
|
Stimulus: Peptides from CSP and AMA1 protein (10 μg/ml) Technic: ELISPOT Process time: 36 h in incubation Cryopreserved: No Supplemented: No |
1 × 106/100 μL/96-well | CSP and AMA1 peptides recalled IFN-γ responses from naturally exposed individuals | Ganeshan et al., 2016 | |
|
Stimulus: TLR1/2 ligand PAM3CSK4 (20 ng) Technic: ND Process time: 72 h in incubation Cryopreserved: Yes Supplemented: ND |
2 × 105/100 μL/96-well | IL-1β and TNF-α were significantly higher in severe malaria cases compared with healthy controls | Manning et al., 2016 | |
|
Stimulus: CelTOS (10 ng) or other single (1.25 μg/ml) peptide pools Technic: ELISPOT Process time: 36 h in incubation Cryopreserved: No Supplemented: No |
4 × 105/100 μL/96-well | Natural malaria transmission induces CelTOS-specific ex vivo IFN-γ | Anum et al., 2015 | |
| Calves PBMC and Cryptosporidium parvum |
Stimulus: Recombinant C. parvum p23 vaccine antigen Technic: Flow Cytometry Process time: ND Cryopreserved: ND Supplemented: ND |
ND | Recombinant p23 vaccine antigen can stimulate a Type-1-like immune response | Wyatt et al., 2005 |
| Human PBMC and Eimeria sp. |
Stimulus: Recombinant Eimeria Antigen (rEA) Technic: Flow Cytometry Process time: 72 h in incubation Cryopreserved: ND Supplemented: 5%FCS |
25 × 106/ND/6-well | rEA stimulates human NK cell effector functions including increasing levels of IFN-γ and Granzyme B | Aylsworth et al., 2013 |
| Sheep PBMC and Babesia sp. |
Stimulus: BdE or BQ1E (10 μg/well) Technic: ELISA Process time: 120 h in incubation Cryopreserved: No Supplemented: 10% autologous plasma |
2 × 105/ND/96-well | Production of IFN-γ and IL10 have key roles in the course of infection by Babesia sp. | Guan et al., 2010 |
| Bovines PBMC and Theileria sp. |
Stimulus: Sporozoites in homogenized infected tick Technic: RT-qPCR Process time: 48 h in incubation Cryopreserved: No Supplemented: 40% FBS |
4 × 106/ND/6-well | MHC-DQ, SIRPA, PRNP, TLR10, cMAF and MAFB genes showed no change in mRNA expression after T. annulata infection | Panigrahi et al., 2016 |
|
Stimulus: Sporozoites in homogenized infected tick Technic: RT-qPCR Process time: 48 h in incubation Cryopreserved: No Supplemented: 40% FBS |
2 × 106/ND/6-well | Up-regulation in SIRPA, PRNP and MHC DQα genes and down-regulation in TLR10, cMAF and MAFB genes in crossbreds as compared to indigenous cattle was observed | Dewangan et al., 2015 | |
|
Stimulus: MPSP Peptides Technic: ELISPOT Process time: 42 h in incubation Cryopreserved: Yes Supplemented: No |
1 × 106/100 μL/96-well | IFN-γ and IL-10 were detected in infected Holsteins but weak responses were exhibited by infected Angus and Japanese Black cattle | Yamaguchi et al., 2010 |
TLA, T. gondii Lysate Antigen; ND, Not determine; OT, Ocular Toxoplasmosis; NC, Negative Control; FCS, Fetal Calf Serum; MPSP, Major piroplasm surface protein; RAMA, Rhoptry-associated membrane antigen; SFC, Spot Forming Cells (SFC)/106; CSP, Plasmodium falciparum circum- sporozoite protein; AMA-1, Apical membrane antigen-1.