Reversal of T cell anergy in leprosy patients: in vitro presentation with Mycobacterium leprae antigens using murabutide and Trat peptide in liposomal delivery

ABSTRACTLsr2 protein ofMycobacterium lepraewas shown earlier to elicit B and T cell responses in leprosy patients (20, 28). Lymphoproliferation toM. lepraeand Lsr2 antigens was observed in >70% of tuberculoid (T) patients and in 16 and 34% of lepromatous (L) patients, respectively. We focused on theM. lepraenonresponders in the lepromatous group using 22 synthetic Lsr2 peptides (end-to-end peptides A to F and overlapping peptides p1 to p16) inin vitroT cell responses. A total of 125 leprosy and 13 tuberculosis patients and 19 healthy controls from the area of endemicity (here, healthy controls, or HC) were investigated. The highest responses were observed (67 to 100%) in HC for all peptides except p1 to p3, and the lowest was observed in tuberculosis patients. Significant differences in lymphoproliferation were observed in T, L, and HC groups (analysis of variance [ANOVA],P= 0.000 to 0.015) for all end-to-end peptides except B and for p5 and p7 to p10. Hierarchical recognition between lepromatous and tuberculoid leprosy was noted for p8 (P< 0.05) and between the HC and L groups for p7 to p10, p15, and p16 (P< 0.005 toP< 0.02). Significant lymphoproliferation was observed to peptides A to F and p1 to p9, p11, p12, p15, p16 (P= 0.000 to 0.001) with 40% responding to peptides C and p16 in L patients. Lepromatous patients also showed significantly higher levels of a gamma interferon (IFN-γ) response to peptide C than to other peptides (P< 0.05). Major histocompatibility complex (MHC) class II bias for peptide recognition was not observed. These studies indicate that Lsr2 has multiple T cell epitopes that inducein vitroT cell responses in the highly infective lepromatous leprosy patients.

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Anti-HLA-DR-triggered monocytes mediate in vitro T cell anergy

International Immunology ◽

10.1093/intimm/dxn019 ◽

2008 ◽

Vol 20 (4) ◽

pp. 601-613 ◽

Cited By ~ 8

Author(s):

Martin A. Kriegel ◽

Sabine Adam-Klages ◽

Christoph Gabler ◽

Norbert Blank ◽

Martin Schiller ◽

...

Keyword(s):

T Cell ◽

T Cell Anergy ◽

Hla Dr ◽

Cell Anergy

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Expansion and Transformation of Primary Acute Lymphoblastic Leukemia Cells into Antigen-Presenting Cells using a Novel Culturing System Enables the Generation of Leukemia-Reactive T Cell Responses that Are Effective in Vivo.

Blood ◽

10.1182/blood.v104.11.303.303 ◽

2004 ◽

Vol 104 (11) ◽

pp. 303-303

Author(s):

Bart A. Nijmeijer ◽

Marianke L.J. Van Schie ◽

Roel Willemze ◽

J.H. Frederik Falkenburg

Keyword(s):

T Cells ◽

T Cell ◽

Cell Lines ◽

Lymphoblastic Leukemia ◽

Primary Cells ◽

T Cell Anergy ◽

Cell Anergy ◽

Antigen Presenting

Abstract Allogeneic cellular immunotherapy is generally ineffective in acute lymphoblastic leukemia (ALL). In vitro studies have suggested that this inefficacy may be the result of a lack of costimulatory molecule expression by ALL cells, resulting in the induction of T cell anergy. Activation of T cells by ALL cells that are transformed into adequate antigen-presenting cells (ALL-APC) may prevent the induction of T cell anergy and result in the generation of competent leukemia-reactive T cell responses for adoptive immunotherapy. However, in vitro modification of ALL cells was hampered by the fact that ALL cells from adult patients could not be cultured in vitro for prolonged periods of time. We have developed a novel serum-free culturing system for B-lineage ALL in which proliferation is initiated and sustained by ALL-cell derived growth factors. Long-term (>2 yrs) proliferation was induced in 12 out of 26 randomly selected primary samples from patients with ALL. The cell cultures ( Leiden cell lines) proliferated with a mean doubling time of 3.0 days (range 2.7–3.6 days). All Leiden cell lines presented the chromosomal abberations observed in the primary cells. The Leiden cell lines displayed an immune phenotype similar to the primary cells, exept for loss of CD34 expression. In vivo characteristics of Leiden cells were evaluated in NOD/scid mice. After intravenous inoculation, Leiden cell lines and primary cells showed identical homing patterns initially involving spleen and bone marrow, followed by the development of overt and progressive leukemia. A comparison of in vivo progression kinetics was performed for one of the Leiden cell lines and the corresponding primary cells. Weekly determination of leukemic cell counts in the blood of engrafted animals revealed that the cell line and the primary cells displayed similar doubling times in vivo of 6.3 and 7.7 days, respectively. To generate cells with improved antigen presentation function, Leiden cell lines were exposed to various activating agents. Stimulation with CpG containing oligonucleotides resulted in induction of CD40 in 9 out of 10 lines. Subsequent ligation of CD40 by culturing CpG-activated Leiden cells on fibroblasts expressing human CD40 ligand resulted in the induction of CD80 or CD86 in 7 of these 10 cell lines. To study the immune stimulatory properties of these Leiden ALL-APC, allogeneic HLA-identical T cells were first activated in vitro by coculturing these cells with either unmodified Leiden cells or with the corresponding Leiden ALL-APC for 3 days, and subsequently infused into groups of 6 leukemic NOD/scid mice. While T cells cocultured with unmodified Leiden cells did not expand in vivo, T cells cocultured with Leiden ALL-APC expanded after infusion in 5 out of 6 animals. This expansion coincided with a 20–75% decrease in leukemic cell numbers in the blood. In conclusion, the novel serum-free culturing system enables long-term culture and manipulation of a significant fraction of primary human ALL. These Leiden cell lines can be modified into ALL-APC that display adequate antigen presenting function, preventing the induction of T cell anergy as demonstrated in vivo in the NOD/scid mouse model.

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Dendritic cells generated with Flt3L and exposed to apoptotic cells lack induction of T cell anergy and Foxp3+ regulatory T cell conversion in vitro

Immunobiology ◽

10.1016/j.imbio.2013.10.006 ◽

2014 ◽

Vol 219 (3) ◽

pp. 230-240 ◽

Cited By ~ 5

Author(s):

Katrien Pletinckx ◽

Manfred B. Lutz

Keyword(s):

Dendritic Cells ◽

T Cell ◽

Regulatory T Cell ◽

Apoptotic Cells ◽

T Cell Anergy ◽

Cell Conversion ◽

Cell Anergy

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Human iPS cell–derived hematopoietic progenitor cells induce T-cell anergy in in vitro–generated alloreactive CD8+ T cells

Blood ◽

10.1182/blood-2012-11-467753 ◽

2013 ◽

Vol 121 (26) ◽

pp. 5167-5175 ◽

Cited By ~ 24

Author(s):

Eun-Mi Kim ◽

Gohar Manzar ◽

Nicholas Zavazava

Keyword(s):

T Cell ◽

Cd8 T Cells ◽

Hematopoietic Progenitor Cells ◽

Hematopoietic Progenitor ◽

Ips Cell ◽

T Cell Anergy ◽

Human Ipscs ◽

Key Points ◽

Cell Anergy

Key Points Human iPSCs differentiate into CD34+ HPCs. iPSC-derived HPCs induce T-cell anergy.

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Cross-reactivity of Mycobacterium leprae and M. tuberculosis and the implications for assessment of in vitro T cell function in leprosy patients

Transactions of the Royal Society of Tropical Medicine and Hygiene ◽

10.1016/0035-9203(89)90294-0 ◽

1989 ◽

Vol 83 (4) ◽

pp. 557-559 ◽

Cited By ~ 1

Author(s):

W.D. Rawlinson ◽

A. Basten

Keyword(s):

T Cell ◽

Cell Function ◽

Mycobacterium Leprae ◽

Cross Reactivity ◽

T Cell Function ◽

Leprosy Patients

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The Avidity Spectrum of T Cell Receptor Interactions Accounts for T Cell Anergy in a Double Transgenic Model

Journal of Experimental Medicine ◽

10.1084/jem.189.2.265 ◽

1999 ◽

Vol 189 (2) ◽

pp. 265-278 ◽

Cited By ~ 38

Author(s):

Laila Girgis ◽

Mark M. Davis ◽

Barbara Fazekas de St. Groth

Keyword(s):

T Cells ◽

Cell Division ◽

T Cell ◽

Cytochrome C ◽

T Cell Receptor ◽

Cytokine Production ◽

Cell Receptor ◽

T Cell Anergy ◽

Cell Anergy

The mechanism of self-tolerance in the CD4+ T cell compartment was examined in a double transgenic (Tg) model in which T cell receptor (TCR)-α/β Tg mice with specificity for the COOH-terminal peptide of moth cytochrome c in association with I-Ek were crossed with antigen Tg mice. Partial deletion of cytochrome-reactive T cells in the thymus allowed some self-specific CD4+ T cells to be selected into the peripheral T cell pool. Upon restimulation with peptide in vitro, these cells upregulated interleukin (IL)-2 receptor but showed substantially lower cytokine production and proliferation than cells from TCR Tg controls. Proliferation and cytokine production were restored to control levels by addition of saturating concentrations of IL-2, consistent with the original in vitro definition of T cell anergy. However, the response of double Tg cells to superantigen stimulation in the absence of exogenous IL-2 was indistinguishable from that of TCR Tg controls, indicating that these self-reactive cells were not intrinsically hyporesponsive. Measurement of surface expression of Tg-encoded TCR α and β chains revealed that cells from double Tg mice expressed the same amount of TCR-β as cells from TCR Tg controls, but only 50% of TCR-α, implying expression of more than one α chain. Naive CD4+ T cells expressing both Tg-encoded and endogenous α chains also manifested an anergic phenotype upon primary stimulation with cytochrome c in vitro, suggesting that low avidity for antigen can produce an anergic phenotype in naive cells. The carboxyfluorescein diacetate succinimidyl ester cell division profiles in response to titered peptide ± IL-2 indicated that expression of IL-2 receptor correlated with peptide concentration but not TCR level, whereas IL-2 production was profoundly affected by the twofold decrease in specific TCR expression. Addition of exogenous IL-2 recruited double Tg cells into division, resulting in a pattern of cell division indistinguishable from that of controls. Thus, in this experimental model, cells expressing more than one α chain escaped negative selection to a soluble self-protein in the thymus and had an anergic phenotype indistinguishable from that of low avidity naive cells. The data are consistent with the notion that avidity-mediated selection for self-reactivity in the thymus may lead to the appearance of anergy within the peripheral, self-reactive T cell repertoire, without invoking the induction of hyporesponsiveness to TCR-mediated signals.

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