Malaria Immunology: Targeting the Surface of Infected Erythrocytes (Methods in Molecular Biology, 2470)

777 Pages·2022·19.061 MB·English

by  Anja Tatiana Ramstedt Jensen (editor), Lars Hviid (editor)

Methods in
Molecular Biology 2470
Anja Tatiana Ramstedt Jensen
Lars Hviid Editors
Malaria
Immunology
Targeting the Surface
of Infected Erythrocytes
M M B
ETHODS IN OLECULAR IO LO GY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire, UK
Forfurther volumes:
http://www.springer.com/series/7651
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Malaria Immunology
Targeting the Surface of Infected Erythrocytes
Edited by
Anja Tatiana Ramstedt Jensen and Lars Hviid
CentreforMedicalParasitology,DepartmentofImmunologyandMicrobiology,UniversityofCopenhagen,
Copenhagen, Denmark
Editors
AnjaTatianaRamstedtJensen LarsHviid
CentreforMedicalParasitology CentreforMedicalParasitology
DepartmentofImmunology DepartmentofImmunology
andMicrobiology andMicrobiology
UniversityofCopenhagen UniversityofCopenhagen
Copenhagen,Denmark Copenhagen,Denmark
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-0716-2188-2 ISBN978-1-0716-2189-9 (eBook)
https://doi.org/10.1007/978-1-0716-2189-9
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Preface
Several books of malaria-related protocols exist—both in general and in the Methods in
MolecularBiologyseries.Incontrasttoitspredecessors,thisbookisfocusedonPlasmodium
falciparum-infectederythrocytes(IEs)andtheirdecisiveroleinasexualblood-stagepathol-
ogy andimmune responses. Ourgoalwastoprovidea systematicand up-to-date one-stop
reference source of protocols to help scientists to understand better how malaria parasite
variantsurfaceantigen-specificpathogenesisandimmunitywork,andtohelpthemtranslate
thatknowledgeintoimprovedinterventionsagainstP.falciparummalaria.ThisMethodsin
Molecular Biology volume includes protocols to support such research all the way from
collecting parasites in the field to single molecule-level analyses of adhesive interactions,
and all the way from basic scientific enquiry to focused studies aiming at disrupting the
devastating disease—the number of casualties of P. falciparum malaria is clearly
unacceptable.
Thevariantsurfaceantigens(VSAs)thattheP.falciparumparasitesputonthesurfaceof
the erythrocytes they infect are the central elements in malaria pathology and immunity.
These molecules, and in particular the P. falciparum erythrocyte membrane protein
1 (PfEMP1) family, are key to the survival of the parasites in vivo and are hence primary
targetsofnaturallyacquiredprotection.Thisparasite-hostconflicthasinevitablyspurredan
evolutionaryarmsraceinvolvingmultipleparasitestrategiestofrustrateandevadedevelop-
mentofprotectivePfEMP1-specificimmunity.Wehopethatthisbookwillhelpresearchers
in their efforts to understand the immune responses and pathology induced by
P.falciparum-IEsandeventuallyachievethegoalsofreducing,eliminating,andeventually
eradicatingmalaria.
Part I contains a series of protocols on how to identify, collect, and preserve
P. falciparum parasites and genetic material in the field. They are followed by protocols
onhowtogettheparasitestogrowininvitrocultures,assesstheirgeneticidentity,studythe
vesiclestheyrelease,andmanipulatethephenotypeandbehavioroftheIEsinvariousways.
Part II presentsseveral protocolson how to assess and controltranscription ofthe var
genesthatencodePfEMP1andoneditingtheP.falciparumgenomebyCRISPR/Cas9.
InPartIII,thefocusisonthePfEMP1proteins.Thesectionincludesprotocolsonthe
purification of native and functionally important PfEMP1molecules, in addition to proto-
cols on expression systems for production of recombinant PfEMP1 and for multiplex
analysisofantibodyresponsestoPfEMP1.
PartIVincludesprotocolsonhowtopurifyPfEMP1-specificantibodiesfromtheblood
of exposed individuals, on the generation of such antibodies by immunization of small
rodents, and regarding production of murine and human monoclonal PfEMP1-specific
antibodies.
PartVcoversassessmentoftheexpressionofVSAsonthesurfaceofP.falciparum-IEs
by a variety of microscopic techniques, as well as protocols for measuring the interaction
betweenPfEMP1andspecifichostmolecules.
Asadhesiontomembrane-boundhostreceptorsisakeyfunctionofPfEMP1,PartVIis
dedicatedtoprotocolsonthatsubject.TheprotocolsincludeELISA-basedassays,staticand
flow-based IE adhesion, adhesion inhibition, and IE selection assays, and assays of the
interaction between IEs with uninfected erythrocytes (rosetting) and platelets. Very recent
v
vi Preface
advances such as IE adhesion to in vitro-engineered microvessels, to self-assembling orga-
noids,andtoPfEMP1-functionalizedlipidmembranesarealsocovered.
Part VII deals with immune effector responses against IEs, other than antibody-
mediated inhibition of IE adhesion covered in the preceding section. It includes protocols
to assess phagocytosis of IEs by monocytes, cytotoxic killing of IEs by neutrophils and
naturalkillercells,andprotocolsfor theevaluationofthecapacityofIE-specificantibodies
toinduceparasiteapoptosisandactivationoftheclassicalcomplementcascade.
The final part (VIII) of the book includes protocols regarding the impact of
P.falciparum infectionexvivoandin vivo.ItcontainsprotocolstoevaluateP. falciparum-
specificBcellfrequenciesandcytokineproductionbycirculatingdendriticcellsinperipheral
bloodsamples,andtoassessdirectlytheinteractionbetweenIEsandendothelialcells—and
itspathologicalconsequences.
Obviously,thisbookwouldbenothingwithoutthecontributionsofthemanyauthors
of its constituent protocols, and we sincerely thank them all for their hard and dedicated
worktomakethiscollectiontakeform.Wealsoverygratefullyacknowledgethenumerous
individuals—somehealthyandsomeill—whoprovidethefoundationofmalariaimmunol-
ogybydonatingsamplesanddatafor research—withouttheseunselfishcontributions,our
researchwouldwither.Ifyoufindtheprotocolsuseful,thecreditistheirs—iferrorsremain,
theresponsibilityisours.Enjoy!
Copenhagen,Denmark AnjaTatianaRamstedtJensen
LarsHviid
Contents
Preface ..................................................................... v
Contributors................................................................. xiii
PART I COLLECTING, GROWING, MONITORING, AND MANIPULATING PLASMODIUM
FALCIPARUM PARASITES
1 IdentificationandEnumerationofPlasmodiumfalciparumParasites
byLightMicroscopy .................................................... 3
AugustinaFrimpong,FredericaDedoPartey,andMichaelFokuoOfori
2 CollectionandCryopreservationofPlasmodiumfalciparumClinical
IsolatesintheField ..................................................... 11
FredericaDedoPartey,AugustinaFrimpong,andMichaelFokuoOfori
3 PreservationofParasiteRNAintheField.................................. 19
BernardTornyigahandNicaiseTuikueNdam
4 PreservationandExtractionofMalariaParasiteDNAfrom
DriedBloodSpots ...................................................... 27
HelleHansson,QueenSaidi,andMichaelAlifrangis
5 EstablishingandMaintainingInVitroCulturesofAsexualBloodStages
ofPlasmodiumfalciparum ............................................... 37
MaryLopez-PerezandZakariaSeidu
6 GenotypingofPlasmodiumfalciparumtoAssessCloneComposition
inParasiteCultures ..................................................... 51
IoannaBroumou,DavidFernandoPlaza,andAnnaFa€rnert
7 ImmunomagneticSelectionofPlasmodiumfalciparum–Infected
ErythrocytesExpressingParticularPfEMP1Variants ........................ 69
MaryLopez-PerezandRebeccaWendelboeOlsen
8 Single-CellSortingofPlasmodiumfalciparum–InfectedErythrocytes
ExpressingParticularPfEMP1Variants.................................... 79
MaryLopez-Perez
9 SelectingPlasmodiumfalciparumInfectedErythrocytesforAdhesion
toCellLines ........................................................... 91
YvonneAzasiandJ.AlexandraRowe
10 SynchronisationofPlasmodiumfalciparumandP.knowlesiInVitro
CulturesUsingaHighlySpecificProteinKinaseInhibitor ................... 101
MargaridaRessurreic¸a˜o,RobertWilliamMoon,DavidAndrewBaker,
andChristiaanvanOoij
11 GenerationofPlasmodiumfalciparumGametocytesInVitro
withSpecificConsiderationsforFieldIsolates.............................. 121
BismarckDinko,RichmondTackie,GiselaHenriques,LindsayStewart,
andColinSutherland
vii
viii Contents
12 ImagingofExtracellularVesiclesDerivedfromPlasmodiumfalciparum–Infected
RedBloodCellsUsingAtomicForceMicroscopy .......................... 133
IritRosenhek-Goldian,PaulaAbouKaram,NetaRegev-Rudzki,and
AliciaRojas
PART II PLASMODIUM FALCIPARUM GENE TRANSCRIPTION AND MANIPULATION
OF PARASITE GENOMES
13 AnalysisofvarGeneTranscriptPatternsbyQuantitative
Real-TimePCR......................................................... 149
AnnaBachmannandThomasLavstsen
14 AnalysisofvarGeneTranscriptionPatternUsingDBLαTags ................ 173
KivisiC.AndisiandAbdirahmanI.Abdi
15 RNAseqofInfectedErythrocyteSurfaceAntigen-EncodingGenes ........... 185
HanhHongThiNguyen,SuffianAzizan,LeeMingYeoh,
JingyiTang,andMichaelF.Duffy
16 ResettingvarGeneTranscriptioninPlasmodiumfalciparum................. 211
VeraMitesserandRonDzikowski
17 CRISPR/Cas9EditingofthePlasmodiumfalciparumGenome.............. 221
SophieAdjalleyandMarcusCheeSanLee
18 CRISPR-Cas9EditingofthePlasmodiumfalciparumGenome:
SpecialApplications..................................................... 241
XuZhang,KirkWilliamDeitsch,andRonDzikowski
PART III PLASMODIUM FALCIPARUM-INFECTED ERYTHROCYTE SURFACE ANTIGENS
19 ExtractionandImmunoprecipitationofVAR2CSA,thePfEMP1
AssociatedwithPlacentalMalaria ......................................... 257
BenoitGamainandDominiqueDorin-Semblat
20 ExpressionofSingle-DomainSolubleandDisulfide-FoldedPfEMP1
AntigensintheEscherichiacoliSHuffleExpressionSystem................... 273
MariadelPilarQuintana
21 ExpressionofLargeFull-LengthPfEMP1ProteinsinHEK293Cells ......... 283
JonathanPaulRenn,JustinYaiAlamouDoritchamou,
andPatrickEmmetDuffy
22 ReceptorAffinity-BasedPurificationofPfEMP1Proteins.................... 299
RebeccaW.Olsen,JenniferSuurbaar,andAnjaRamstedtJensen
23 AnalysisofAntibodyReactivitytoMalariaAntigensbyMicrosphere-Based
MultiplexImmunoassay ................................................. 309
IsobelS.Walker,AmyW.Chung,TimonDamelang,andStephenJ.Rogerson
24 High-ThroughputBioPlexAssayfor theStudyofFunctionally
ActivePlasmodiumFalciparumAntigensThatAreExpressed
ontheSurfaceofInfectedErythrocytes.................................... 327
AndrewV.Oleinikov
Contents ix
25 ProteinMicroarraysasaTooltoAnalyzeAntibodyResponses
toVariantSurfaceAntigensExpressedontheSurfaceofPlasmodium
falciparum–InfectedErythrocytes......................................... 343
AlbertE.Zhou,AartiJain,RieNakajima,BirajShrestha,EmilyM.Stucke,
SudhaunshuJoshi,KathyA.Strauss,PerN.Hedde,AndreaA.Berry,
PhilipL.Felgner,andMarkA.Travassos
26 ProtocolforDifferentialBiopanningofP.falciparumPhageDisplaycDNA
LibrarytoIdentifyParasiteTargetsofProtectiveAntibodies ................. 359
JennaZuromski,JonathanKurtis,andDipakKumarRaj
PART IV ANTIBODIES TO PLASMODIUM FALCIPARUM-INFECTED
ERYTHROCYTE SURFACE ANTIGENS
27 AffinityPurificationofPfEMP1-SpecificAntibodies
fromHumanBlood..................................................... 369
RebeccaW.Olsen,JenniferSuurbaar,andAnjaRamstedtJensen
28 Productionofanti-PfEMP1PolyclonalAntiserainRatsandMice ............ 381
RebeccaW.Olsen,JenniferSuurbaar,andAnjaRamstedtJensen
29 ProductionofPfEMP1-SpecificMouseMonoclonalAntibodies.............. 391
NannaDalgaardandLeaBarfod
30 ProductionofPfEMP1-SpecificHumanMonoclonalAntibodies
fromNaturallyImmuneIndividuals....................................... 407
MelanieR.WalkerandLeaBarfod
PART V ANALYSIS OF ANTIGENS ON PLASMODIUM
FALCIPARUM-INFECTED ERYTHROCYTES
31 VisualizationofInfectedRedBloodCellSurfaceAntigensbyFluorescence
Microscopy ............................................................ 425
MarkusGanter,JulienGuizetti,andNicoleKilian
32 AnalysisbyFlowCytometryofα -MacroglobulinandNonimmune
2
IgM-BindingtoPlasmodiumfalciparum-InfectedErythrocytes .............. 435
LarsHviidandMaryLopez-Perez
33 AnImprovedMethodforAssessingAntigenPresentationontheSurface
ofPlasmodiumfalciparum-InfectedErythrocytes
byImmuno-ElectronMicroscopy......................................... 445
MarekCyrklaff,CeciliaPalmiraSanchez,LukasHanebutte,JuliaJa€ger,
UlrichS.Schwarz,andMichaelLanzer
34 AssessingAntigenPresentationontheSurfaceofPlasmodium
falciparum-InfectedErythrocytesbyPhotoactivatedLocalization
Microscopy(PALM) .................................................... 457
ChristosKarathanasis,CeciliaP.Sanchez,MikeHeilemann,
andMichaelLanzer
35 SurfacePlasmonResonanceAnalysisofPfEMP1
InteractionwithReceptors............................................... 467
FrankLennartzandMatthewK.Higgins