Human Antibodies against Plasmodium falciparumLiver-Stage Antigen 3 Cross-React with Plasmodium yoelii Preerythrocytic-Stage Epitopes and Inhibit Sporozoite Invasion In Vitro and In Vivo

ABSTRACT The Plasmodium falciparum liver-stage antigen 3 (LSA3), a recently identified preerythrocytic antigen, induces protection against malaria in chimpanzees. Using antibodies from individuals with hyperimmunity to malaria affinity purified on recombinant or synthetic polypeptides of LSA3, we identified four non-cross-reactive B-cell epitopes in Plasmodium yoelii preerythrocytic stages. On sporozoites the P. yoelii protein detected has a molecular mass similar to that of LSA3. T-cell epitopes cross-reacting withP. yoelii were also demonstrated using peripheral blood lymphocytes from LSA3-immunized chimpanzees. In contrast, no cross-reactive epitopes were found in Plasmodium berghei. LSA3-specific human antibodies exerted up to 100% inhibition of in vitro invasion of P. yoelii sporozoites into mouse hepatocytes. This strong in vitro activity was reproduced in vivo by passive transfer of LSA3 antibodies. These results indicate that the homologous epitopes may be biologically functional and suggest that P. yoelii could be used as a model to assess the antisporozoite activity of anti-LSA3 antibodies.

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Yellow fever 17D as a vaccine vector for microbial CTL epitopes

Journal of Experimental Medicine ◽

10.1084/jem.20041526 ◽

2005 ◽

Vol 201 (2) ◽

pp. 201-209 ◽

Cited By ~ 49

Author(s):

Deng Tao ◽

Giovanna Barba-Spaeth ◽

Urvashi Rai ◽

Victor Nussenzweig ◽

Charles M. Rice ◽

...

Keyword(s):

T Cell ◽

Yellow Fever ◽

Cell Epitope ◽

Parasite Burden ◽

Plasmodium Yoelii ◽

Yellow Fever Vaccine ◽

T Cell Epitopes ◽

High Level

The yellow fever vaccine 17D (17D) is safe, and after a single immunizing dose, elicits long-lasting, perhaps lifelong protective immunity. One of the major challenges facing delivery of human vaccines in underdeveloped countries is the need for multiple injections to achieve full efficacy. To examine 17D as a vector for microbial T cell epitopes, we inserted the H-2Kd–restricted CTL epitope of the circumsporozoite protein (CS) of Plasmodium yoelii between 17D nonstructural proteins NS2B and NS3. The recombinant virus, 17D-Py, was replication competent and stable in vitro and in vivo. A single subcutaneous injection of 105 PFU diminished the parasite burden in the liver by ∼70%. The high level of protection lasted between 4 and 8 wk after immunization, but a significant effect was documented even 24 wk afterwards. Thus, the immunogenicity of a foreign T cell epitope inserted into 17D mimics some of the remarkable properties of the human vaccine. Priming with 17D-Py followed by boosting with irradiated sporozoites conferred sterile immunity to 90% of the mice. This finding indicates that the immune response of vaccine-primed individuals living in endemic areas could be sustained and magnified by the bite of infected mosquitoes.

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Identification of Plasmodium falciparum proteoforms from liver stage models

Malaria Journal ◽

10.1186/s12936-019-3093-3 ◽

2020 ◽

Vol 19 (1) ◽

Author(s):

Benjamin Winer ◽

Kimberly A. Edgel ◽

Xiaoyan Zou ◽

Julie Sellau ◽

Sri Hadiwidjojo ◽

...

Keyword(s):

Mass Spectrometry ◽

Plasmodium Falciparum ◽

Vaccine Development ◽

Strongly Correlated ◽

T Cell Epitopes ◽

Liver Stage ◽

Stage Models ◽

Erythrocyte Antigen

Abstract Background Immunization with attenuated malaria sporozoites protects humans from experimental malaria challenge by mosquito bite. Protection in humans is strongly correlated with the production of T cells targeting a heterogeneous population of pre-erythrocyte antigen proteoforms, including liver stage antigens. Currently, few T cell epitopes derived from Plasmodium falciparum, the major aetiologic agent of malaria in humans are known. Methods In this study both in vitro and in vivo malaria liver stage models were used to sequence host and pathogen proteoforms. Proteoforms from these diverse models were subjected to mild acid elution (of soluble forms), multi-dimensional fractionation, tandem mass spectrometry, and top-down bioinformatics analysis to identify proteoforms in their intact state. Results These results identify a group of host and malaria liver stage proteoforms that meet a 5% false discovery rate threshold. Conclusions This work provides proof-of-concept for the validity of this mass spectrometry/bioinformatic approach for future studies seeking to reveal malaria liver stage antigens towards vaccine development.

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Identification of an HLA-A*0201-restricted cytotoxic T-lymphocyte epitope in rotavirus VP6 protein

Journal of General Virology ◽

10.1099/vir.0.82031-0 ◽

2006 ◽

Vol 87 (11) ◽

pp. 3393-3396 ◽

Cited By ~ 9

Author(s):

Jing Wei ◽

Jin-Tao Li ◽

Xiao-Ping Zhang ◽

Yan Tang ◽

Jing-Xue Wang ◽

...

Keyword(s):

Cytotoxic T Lymphocyte ◽

Human Leukocyte ◽

Hla Class I ◽

Peripheral Blood Lymphocytes ◽

T Cell Epitopes ◽

Computer Algorithms ◽

Leukocyte Antigen ◽

Healthy Donors

The function of cytotoxic T lymphocytes (CTLs) in rotavirus (RV) infection in humans is poorly understood. To date, no RV-specific human leukocyte antigen (HLA) class I-restricted T-cell epitopes have been described. In this study, four peptides derived from human RV Wa strain VP6 protein were predicted by computer algorithms and verified by an HLA*0201-binding assay. Two peptides with high affinity for HLA-A*0201 molecules were further assessed. The CTLs induced in vitro by P340–348 (TLLANVTAV)-loaded autologous dendritic cells from peripheral blood lymphocytes of HLA-A*0201-matched healthy donors released gamma interferon specifically upon stimulation with P340–348-loaded T2 cells. The CTLs lysed both P340–348-loaded T2 cells and human RV Wa strain-infected HLA-A*0201+ Caco-2 cells in an antigen-specific and HLA-A*0201-restricted manner. At the same time, P340–348 was shown to be immunogenic in vivo in HLA-A*0201/Kb transgenic mice. It is proposed that P340–348 is an HLA-A*0201-restricted CTL epitope.

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Mouse models to study von Willebrand factor structure-function relationships in vivo

Hämostaseologie ◽

10.1055/s-0037-1616933 ◽

2009 ◽

Vol 29 (01) ◽

pp. 17-20 ◽

Cited By ~ 2

Author(s):

I. Marx ◽

I. Badirou ◽

R. Pendu ◽

O. Christophe ◽

C. V. Denis

Keyword(s):

Structure Function ◽

Transient Expression ◽

Von Willebrand Factor ◽

Large Size ◽

Mouse Hepatocytes ◽

Function Relationship ◽

Von Willebrand ◽

Willebrand Factor

SummaryVon Willebrand factor (VWF) structure-function relationship has been studied only through in vitro approaches. The VWF-deficient mouse model has been extremely useful to examine the in vivo function of VWF but does not allow a more subtle analysis of the relative importance of its different domains. However, considering the large size of VWF and its capacity to interact with various ligands in order to support platelet adhesion and aggregation, the necessity to evaluate independently these interactions appeared increasingly crucial. A recently developed technique, known as hydrodynamic injection, which allows transient expression of a transgene by mouse hepatocytes, proved very useful in this regard. Indeed, transient expression of various VWF mutants in VWF-deficient mice contributed to improve our knowledge about the role of VWF interaction with subendothelial collagens and with platelets receptors in VWF roles in haemostasis and thrombosis. These findings can provide new leads in the development of anti-thrombotic therapies.

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Allergenic Characterization of New Mutant Forms of Pru p 3 as New Immunotherapy Vaccines

Clinical and Developmental Immunology ◽

10.1155/2013/385615 ◽

2013 ◽

Vol 2013 ◽

pp. 1-12 ◽

Cited By ~ 6

Author(s):

C. Gómez-Casado ◽

M. Garrido-Arandia ◽

P. Gamboa ◽

N. Blanca-López ◽

G. Canto ◽

...

Keyword(s):

Food Allergy ◽

Ex Vivo ◽

Binding Capacity ◽

Cross Reactivity ◽

T Cell Epitopes ◽

Native Form ◽

Ige Binding ◽

Pru P 3

Nowadays, treatment of food allergy only considered the avoidance of the specific food. However, the possibility of cross-reactivity makes this practice not very effective. Immunotherapy may exhibit as a good alternative to food allergy treatment. The use of hypoallergenic molecules with reduced IgE binding capacity but with ability to stimulate the immune system is a promising tool which could be developed for immunotherapy. In this study, three mutants of Pru p 3, the principal allergen of peach, were produced based on the described mimotope and T cell epitopes, by changing the specific residues to alanine, named asPru p 3.01, Pru p 3.02, andPru p 3.03.Pru p 3.01showed very similar allergenic activity as the wild type byin vitroassays. However,Pru p 3.02andPru p 3.03presented reduced IgE binding with respect to the native form, byin vitro,ex vivo,and in vivo assays. In addition,Pru p 3.03had affected the IgG4 binding capacity and presented a random circular dichroism, which was reflected in the nonrecognition by specific antibodies anti-Pru p 3. Nevertheless, bothPru p 3.02andPru p 3.03maintained the binding to IgG1 and their ability to activate T lymphocytes. Thus,Pru p 3.02andPru p 3.03could be good candidates for potential immunotherapy in peach-allergic patients.

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NUANCE, a giant protein connecting the nucleus and actin cytoskeleton

Journal of Cell Science ◽

10.1242/jcs.115.15.3207 ◽

2002 ◽

Vol 115 (15) ◽

pp. 3207-3222 ◽

Cited By ~ 25

Author(s):

Yen-Yi Zhen ◽

Thorsten Libotte ◽

Martina Munck ◽

Angelika A. Noegel ◽

Elena Korenbaum

Keyword(s):

Actin Cytoskeleton ◽

Transmembrane Domain ◽

Coiled Coil ◽

Peripheral Blood Lymphocytes ◽

Binding Domain ◽

Actin Binding ◽

Actin Binding Domain ◽

Microfilament System

NUANCE (NUcleus and ActiN Connecting Element) was identified as a novel protein with an α-actinin-like actin-binding domain. A human 21.8 kb cDNA of NUANCE spreads over 373 kb on chromosome 14q22.1-q22.3. The cDNA sequence predicts a 796 kDa protein with an N-terminal actin-binding domain, a central coiled-coil rod domain and a predicted C-terminal transmembrane domain. High levels of NUANCE mRNA were detected in the kidney, liver,stomach, placenta, spleen, lymphatic nodes and peripheral blood lymphocytes. At the subcellular level NUANCE is present predominantly at the outer nuclear membrane and in the nucleoplasm. Domain analysis shows that the actin-binding domain binds to Factin in vitro and colocalizes with the actin cytoskeleton in vivo as a GFP-fusion protein. The C-terminal transmembrane domain is responsible for the targeting the nuclear envelope. Thus, NUANCE is the firstα-actinin-related protein that has the potential to link the microfilament system with the nucleus.

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Interferon beta-1a induces expression of brain-derived neurotrophic factor in human T lymphocytes in vitro and not in vivo

Future Neurology ◽

10.2217/fnl-2019-0018 ◽

2020 ◽

Vol 15 (1) ◽

pp. FNL38 ◽

Cited By ~ 1

Author(s):

Zarlascht Karmand ◽

Hans-Peter Hartung ◽

Oliver Neuhaus

Keyword(s):

T Cell ◽

Neurotrophic Factor ◽

Serum Levels ◽

Brain Derived Neurotrophic Factor ◽

Peripheral Blood Lymphocytes ◽

T Cell Proliferation ◽

Bdnf Expression ◽

Healthy Donors

Aim: To detect IFN β-1a-induced expression of brain-derived neurotrophic factor (BDNF) to undermine the hypothesis of IFN β-1a-associated neuroprotection in multiple sclerosis (MS). Methods: The influence of IFN β-1a on in vitro activated peripheral blood lymphocytes from healthy donors was tested. Proliferation analyses were made to detect T-cell growth. BDNF expression was measured by standard ELISA. To assess the influence of IFN β-1a on BDNF expression in vivo, BDNF serum levels of MS patients treated with IFN β-1a were compared with those of untreated patients. Results: IFN β-1a inhibited T-cell proliferation dose dependently. It induced BDNF expression at middle concentrations. MS patients treated with IFN β-1a exhibited significantly lower BDNF serum levels than untreated patients. Conclusion: IFN β-1a may promote neuroprotection by inducing BDNF expression, but its importance in vivo remains open.

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Plasmodium yoelii: Efficient in Vitro Invasion and Complete Development of Sporozoites in Mouse Hepatic Cell Lines

Experimental Parasitology ◽

10.1006/expr.2000.4570 ◽

2000 ◽

Vol 96 (4) ◽

pp. 257-259 ◽

Cited By ~ 18

Author(s):

Maria M Mota ◽

Ana Rodriguez

Keyword(s):

Cell Lines ◽

Plasmodium Yoelii ◽

Hepatic Cell ◽

Complete Development ◽

In Vitro Invasion ◽

Hepatic Cell Lines

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Interleukin 13 inhibits human immunodeficiency virus type 1 production in primary blood-derived human macrophages in vitro.

Journal of Experimental Medicine ◽

10.1084/jem.178.2.743 ◽

1993 ◽

Vol 178 (2) ◽

pp. 743-747 ◽

Cited By ~ 79

Author(s):

L J Montaner ◽

A G Doyle ◽

M Collin ◽

G Herbein ◽

P Illei ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Dna Analysis ◽

Peripheral Blood Lymphocytes ◽

Interleukin 13 ◽

Immunodeficiency Virus

The mechanisms by which cellular immunity maintains the asymptomatic state after human immunodeficiency virus type 1 (HIV-1) infection are poorly understood. CD4+ T lymphocytes play a complex role in regulating anti-HIV effector pathways, including activation of macrophages, which are themselves implicated in clinical latency and pathogenesis of symptomatic acquired immune deficiency syndrome. We have found that a newly identified T helper type 2 lymphokine, interleukin 13 (IL-13), inhibits HIV-1ADA and Ba-L replication in primary tissue culture-derived macrophages but not in peripheral blood lymphocytes. Viral production in cells was measured by viral protein (p24) and reverse transcriptase levels, while entry was assessed by proviral DNA analysis at timed intervals after infection. Inhibition by IL-13 was dose and time dependent and not mediated through altered viral entry, reverse transcription, or viral release. IL-13 is therefore a candidate cytokine for the suppression of HIV infection within monocytes and macrophages in vivo.

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