Usage¶
Warning
Using a local IEDB installation is strongly recommended for larger datasets or when the making predictions for many alleles, epitope lengths, or prediction algorithms. More information on how to install IEDB locally can be found on the Installation page.
It may be necessary to explore the parameter space a bit when running pVACvector. As binding predictions for some sites vary substantially across algorithms, the most conservative settings may result in no valid paths, often due to one “outlier” prediction. Carefully choosing which predictors to run may help ameliorate this issue as well.
In general, setting a lower binding threshold (e.g., 500nM) and using the median
binding value (--top-score-metric median) will lead to greater possibility
of a design, while more conservative settings of 1000nM and lowest/best binding
value (--top-score-metric lowest) will give more confidence that there are
no junctional neoepitopes.
Our current recommendation is to run pVACvector several different ways, and choose the path resulting from the most conservative set of parameters.
usage: pvacvector run [-h] [-e1 CLASS_I_EPITOPE_LENGTH]
[-e2 CLASS_II_EPITOPE_LENGTH]
[--iedb-install-directory IEDB_INSTALL_DIRECTORY]
[-b BINDING_THRESHOLD]
[--percentile-threshold PERCENTILE_THRESHOLD]
[--allele-specific-binding-thresholds]
[--aggregate-inclusion-binding-threshold AGGREGATE_INCLUSION_BINDING_THRESHOLD]
[-m {lowest,median}] [-r IEDB_RETRIES] [-k]
[-t N_THREADS] [-v INPUT_VCF] [-n INPUT_N_MER]
[--spacers SPACERS] [--max-clip-length MAX_CLIP_LENGTH]
input_file sample_name allele
{BigMHC_EL,BigMHC_IM,DeepImmuno,MHCflurry,MHCflurryEL,MHCnuggetsI,MHCnuggetsII,NNalign,NetMHC,NetMHCIIpan,NetMHCIIpanEL,NetMHCcons,NetMHCpan,NetMHCpanEL,PickPocket,SMM,SMMPMBEC,SMMalign,all,all_class_i,all_class_ii}
[{BigMHC_EL,BigMHC_IM,DeepImmuno,MHCflurry,MHCflurryEL,MHCnuggetsI,MHCnuggetsII,NNalign,NetMHC,NetMHCIIpan,NetMHCIIpanEL,NetMHCcons,NetMHCpan,NetMHCpanEL,PickPocket,SMM,SMMPMBEC,SMMalign,all,all_class_i,all_class_ii} ...]
output_dir
Run the pVACvector pipeline
positional arguments:
input_file A .fa file with peptides or a pVACseq .tsv file with
epitopes to use for vector design.
sample_name The name of the sample being processed. This will be
used as a prefix for output files.
allele Name of the allele to use for epitope prediction.
Multiple alleles can be specified using a comma-
separated list. For a list of available alleles, use:
`pvacvector valid_alleles`.
{BigMHC_EL,BigMHC_IM,DeepImmuno,MHCflurry,MHCflurryEL,MHCnuggetsI,MHCnuggetsII,NNalign,NetMHC,NetMHCIIpan,NetMHCIIpanEL,NetMHCcons,NetMHCpan,NetMHCpanEL,PickPocket,SMM,SMMPMBEC,SMMalign,all,all_class_i,all_class_ii}
The epitope prediction algorithms to use. Multiple
prediction algorithms can be specified, separated by
spaces.
output_dir The directory for writing all result files.
optional arguments:
-h, --help show this help message and exit
-e1 CLASS_I_EPITOPE_LENGTH, --class-i-epitope-length CLASS_I_EPITOPE_LENGTH
Length of MHC Class I subpeptides (neoepitopes) to
predict. Multiple epitope lengths can be specified
using a comma-separated list. Typical epitope lengths
vary between 8-15. Required for Class I prediction
algorithms. (default: [8, 9, 10, 11])
-e2 CLASS_II_EPITOPE_LENGTH, --class-ii-epitope-length CLASS_II_EPITOPE_LENGTH
Length of MHC Class II subpeptides (neoepitopes) to
predict. Multiple epitope lengths can be specified
using a comma-separated list. Typical epitope lengths
vary between 11-30. Required for Class II prediction
algorithms. (default: [12, 13, 14, 15, 16, 17, 18])
--iedb-install-directory IEDB_INSTALL_DIRECTORY
Directory that contains the local installation of IEDB
MHC I and/or MHC II. (default: None)
-b BINDING_THRESHOLD, --binding-threshold BINDING_THRESHOLD
Report only epitopes where the mutant allele has ic50
binding scores below this value. (default: 500)
--percentile-threshold PERCENTILE_THRESHOLD
Report only epitopes where the mutant allele has a
percentile rank below this value. (default: None)
--allele-specific-binding-thresholds
Use allele-specific binding thresholds. To print the
allele-specific binding thresholds run `pvacvector
allele_specific_cutoffs`. If an allele does not have a
special threshold value, the `--binding-threshold`
value will be used. (default: False)
--aggregate-inclusion-binding-threshold AGGREGATE_INCLUSION_BINDING_THRESHOLD
Threshold for including epitopes when creating the
aggregate report (default: 5000)
-m {lowest,median}, --top-score-metric {lowest,median}
The ic50 scoring metric to use when filtering epitopes
by binding-threshold or minimum fold change. lowest:
Use the best MT Score and Corresponding Fold Change
(i.e. the lowest MT ic50 binding score and
corresponding fold change of all chosen prediction
methods). median: Use the median MT Score and Median
Fold Change (i.e. the median MT ic50 binding score and
fold change of all chosen prediction methods).
(default: median)
-r IEDB_RETRIES, --iedb-retries IEDB_RETRIES
Number of retries when making requests to the IEDB
RESTful web interface. Must be less than or equal to
100. (default: 5)
-k, --keep-tmp-files Keep intermediate output files. This might be useful
for debugging purposes. (default: False)
-t N_THREADS, --n-threads N_THREADS
Number of threads to use for parallelizing peptide-MHC
binding prediction calls. (default: 1)
-v INPUT_VCF, --input_vcf INPUT_VCF
Path to original pVACseq input VCF file. Required if
input file is a pVACseq TSV. (default: None)
-n INPUT_N_MER, --input-n-mer INPUT_N_MER
Length of the peptide sequence to use when creating
the FASTA from the pVACseq TSV. (default: 25)
--spacers SPACERS Comma-separated list of spacers to use for testing
junction epitopes. Include None to test junctions
without spacers. Peptide combinations will be tested
with each spacer in the order specified. (default: Non
e,AAY,HHHH,GGS,GPGPG,HHAA,AAL,HH,HHC,HHH,HHHD,HHL,HHHC
)
--max-clip-length MAX_CLIP_LENGTH
Number of amino acids to permit clipping from the
start and/or end of peptides in order to test novel
junction epitopes when the first pass on the full
peptide fails. (default: 3)