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.

usage: pvacseq run [-h] [-e EPITOPE_LENGTH] [-l PEPTIDE_SEQUENCE_LENGTH]
                   [--iedb-install-directory IEDB_INSTALL_DIRECTORY]
                   [-i ADDITIONAL_INPUT_FILE_LIST]
                   [--net-chop-method {cterm,20s}] [--netmhc-stab] [-t]
                   [-m {lowest,median}] [-b BINDING_THRESHOLD]
                   [-c MINIMUM_FOLD_CHANGE] [--normal-cov NORMAL_COV]
                   [--tdna-cov TDNA_COV] [--trna-cov TRNA_COV]
                   [--normal-vaf NORMAL_VAF] [--tdna-vaf TDNA_VAF]
                   [--trna-vaf TRNA_VAF] [--expn-val EXPN_VAL]
                   [--net-chop-threshold NET_CHOP_THRESHOLD]
                   [-a {sample_name}] [-s FASTA_SIZE] [-r IEDB_RETRIES]
                   [-d DOWNSTREAM_SEQUENCE_LENGTH] [-k]
                   input_file sample_name allele
                   {NNalign,NetMHC,NetMHCIIpan,NetMHCcons,NetMHCpan,PickPocket,SMM,SMMPMBEC,SMMalign}
                   [{NNalign,NetMHC,NetMHCIIpan,NetMHCcons,NetMHCpan,PickPocket,SMM,SMMPMBEC,SMMalign} ...]
                   output_dir

Positional Arguments

input_file A VEP-annotated single-sample VCF containing transcript, Wildtype protein sequence, and Downstream protein sequence information
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: pvacseq valid_alleles
prediction_algorithms
 

Possible choices: NNalign, NetMHC, NetMHCIIpan, NetMHCcons, NetMHCpan, PickPocket, SMM, SMMPMBEC, SMMalign

The epitope prediction algorithms to use. Multiple prediction algorithms can be specified, separated by spaces

output_dir The directory for writing all result files

Named Arguments

-e, –epitope-length
 Length of subpeptides (neoepitopes) to predict. Multiple epitope lengths can be specified using a comma-separated list. Typical epitope lengths vary between 8-11. Required for Class I prediction algorithms
-l, –peptide-sequence-length
 

Length of the peptide sequence to use when creating the FASTA. Default: 21

Default: 21

–iedb-install-directory
 Directory that contains the local installation of IEDB MHC I and/or MHC II
-i, –additional-input-file-list
 yaml file of additional files to be used as inputs, e.g. cufflinks output files. For an example of this yaml file run pvacseq config_files additional_input_file_list.
–net-chop-method
 

Possible choices: cterm, 20s

NetChop prediction method to use (“cterm” for C term 3.0, “20s” for 20S 3.0).

–netmhc-stab

Run NetMHCStabPan after all filtering and add stability predictions to predicted epitopes

Default: False

-t, –top-result-per-mutation
 

Output only the top scoring result for each allele-peptide length combination for each variant. Default: False

Default: False

-m, –top-score-metric
 

Possible choices: lowest, median

The ic50 scoring metric to use when filtering epitopes by binding-threshold or minimum fold change. lowest: Best MT Score/Corresponding Fold Change - lowest MT ic50 binding score/corresponding fold change of all chosen prediction methods. median: Median MT Score/Median Fold Change - median MT ic50 binding score/fold change of all chosen prediction methods. Default: median

Default: “median”

-b, –binding-threshold
 

Report only epitopes where the mutant allele has ic50 binding scores below this value. Default: 500

Default: 500

-c, –minimum-fold-change
 

Minimum fold change between mutant binding score and wild-type score. The default is 0, which filters no results, but 1 is often a sensible choice (requiring that binding is better to the MT than WT). Default: 0

Default: 0

–normal-cov

Normal Coverage Cutoff. Sites above this cutoff will be considered. Default: 5

Default: 5

–tdna-cov

Tumor DNA Coverage Cutoff. Sites above this cutoff will be considered. Default: 10

Default: 10

–trna-cov

Tumor RNA Coverage Cutoff. Sites above this cutoff will be considered. Default: 10

Default: 10

–normal-vaf

Normal VAF Cutoff. Sites BELOW this cutoff in normal will be considered. Default: 2

Default: 2

–tdna-vaf

Tumor DNA VAF Cutoff. Sites above this cutoff will be considered. Default: 40

Default: 40

–trna-vaf

Tumor RNA VAF Cutoff. Sites above this cutoff will be considered. Default: 40

Default: 40

–expn-val

Gene and Transcript Expression cutoff. Sites above this cutoff will be considered. Default: 1

Default: 1

–net-chop-threshold
 

NetChop prediction threshold. Default: 0.5

Default: 0.5

-a, –additional-report-columns
 

Possible choices: sample_name

Additional columns to output in the final report

-s, –fasta-size
 

Number of fasta entries per IEDB request. For some resource-intensive prediction algorithms like Pickpocket and NetMHCpan it might be helpful to reduce this number. Needs to be an even number.

Default: 200

-r, –iedb-retries
 

Number of retries when making requests to the IEDB RESTful web interface. Must be less than or equal to 100.Default: 5

Default: 5

-d, –downstream-sequence-length
 

Cap to limit the downstream sequence length for frameshifts when creating the fasta file. Use ‘full’ to include the full downstream sequence. Default: 1000

Default: “1000”

-k, –keep-tmp-files
 

Keep intermediate output files. This migt be useful for debugging purposes.

Default: False